NCERT - BIOTECHNOLOGY - PRINCIPLES & PROCESSES

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Questions and Answers

What significant property do restriction enzymes discovered by Boyer possess?

They can replicate DNA.

They can separate plasmids from bacterial cells.

They can cut DNA strands leaving 'sticky ends'. (correct)

They can transform one organism into another.

Herbert Boyer completed his undergraduate studies at Yale.

False

Who collaborated with Herbert Boyer in the early development of biotechnology?

Stanley Cohen

Biotechnology uses live organisms or __________ from organisms to produce useful products.

enzymes Signup and view all the answers

Match the following terms with their correct descriptions:

Restriction enzymes = Cut DNA to create 'sticky ends' Plasmids = Small ringlets of DNA in bacterial cells Biotechnology = Using organisms to create products Recombinant DNA = DNA formed by splicing together segments of DNA Signup and view all the answers

What process did Boyer and Cohen enable bacteria to perform?

Manufacture specific proteins Signup and view all the answers

Making bread could be considered a form of biotechnology.

True Signup and view all the answers

In what year did Herbert Boyer become an assistant professor at the University of California at San Francisco?

1966 Signup and view all the answers

Which of the following techniques is primarily associated with altering the chemistry of genetic material?

Genetic engineering Signup and view all the answers

Asexual reproduction provides more genetic variation than sexual reproduction.

False Signup and view all the answers

What is the main advantage of using genetic engineering over traditional hybridization methods?

It allows isolation and introduction of desirable genes without including undesirable genes. Signup and view all the answers

Bioprocess engineering maintains a sterile ambiance to enable growth of only the desired ______ in large quantities.

microbe Signup and view all the answers

Match the following terms with their corresponding descriptions:

Genetic engineering = Techniques to alter DNA and RNA Bioprocess engineering = Maintaining sterile conditions for microbial growth Recombinant DNA = DNA that has been artificially made using two different organisms Hybridization = Mating of organisms to produce offspring with desired traits Signup and view all the answers

Which organization provided a definition for biotechnology?

European Federation of Biotechnology Signup and view all the answers

Incorporating a piece of DNA into an alien organism guarantees its multiplication in progeny cells.

False Signup and view all the answers

What is the significant impact of sexual reproduction compared to asexual reproduction on genetic traits?

Sexual reproduction allows the introduction of variations and beneficial genetic combinations. Signup and view all the answers

Which of the following is the first step in genetically modifying an organism?

Identification of DNA with desirable genes Signup and view all the answers

Restriction enzymes only make cuts at the ends of DNA molecules.

False Signup and view all the answers

What is the significance of the recognition sequence in restriction enzymes?

It is the specific sequence of base pairs where the restriction enzyme cuts the DNA. Signup and view all the answers

The first restriction endonuclease, Hind II, recognizes a specific sequence of _____ base pairs.

six Signup and view all the answers

Match the following restriction enzymes with their origin:

Hind II = Haemophilus influenzae EcoRI = Escherichia coli BglII = Bacillus globigii PstI = Providencia stuartii Signup and view all the answers

Which of the following enzymes is responsible for cutting DNA?

Restriction endonucleases Signup and view all the answers

Exonucleases can only remove nucleotides from the ends of the DNA.

True Signup and view all the answers

A major tool in recombinant DNA technology that introduces DNA into the host is called a _____ .

vector Signup and view all the answers

What do restriction endonucleases cut in the DNA?

The sugar-phosphate backbone Signup and view all the answers

Restriction enzymes can create blunt ends when they cut DNA.

False Signup and view all the answers

What are the single-stranded portions at the ends of a cut DNA called?

sticky ends Signup and view all the answers

Restriction endonucleases recognize specific sequences known as __________.

palindromic nucleotide sequences Signup and view all the answers

Match the following terms with their definitions:

Restriction Endonuclease = Enzyme that cuts DNA at specific sequences Sticky Ends = Single-stranded overhanging portions after cutting Recombinant DNA = DNA formed from different sources DNA Ligase = Enzyme that joins DNA fragments together Signup and view all the answers

Which of the following statements about palindromic DNA sequences is true?

They form the same sequence when read in both directions. Signup and view all the answers

Different restriction enzymes can create compatible sticky ends.

True Signup and view all the answers

When two DNA fragments are cut using the same restriction enzyme, they can be joined together by __________.

DNA ligase Signup and view all the answers

What is the primary component used in gel electrophoresis as the matrix?

Agarose Signup and view all the answers

DNA fragments can be visualized with the naked eye without staining.

False Signup and view all the answers

What color do stained DNA bands appear after exposure to UV light?

Bright orange Signup and view all the answers

In gel electrophoresis, DNA fragments migrate towards the ______ because they are negatively charged.

anode Signup and view all the answers

Which of the following correctly describes the movement of DNA fragments during gel electrophoresis?

Smaller fragments move farther than larger fragments. Signup and view all the answers

Elution involves cutting the DNA bands from the agarose gel.

True Signup and view all the answers

What is the purpose of using ethidium bromide in gel electrophoresis?

To visualize DNA fragments Signup and view all the answers

What is a primary function of the origin of replication (ori) in a cloning vector?

To initiate replication of linked DNA Signup and view all the answers

Bacteriophages have a lower copy number of their genome compared to plasmids within bacterial cells.

False Signup and view all the answers

What is the significance of having a selectable marker in a cloning vector?

It helps in identifying and eliminating non-transformants. Signup and view all the answers

The genes encoding resistance to antibiotics such as ampicillin and __________ are commonly used as selectable markers in E. coli.

tetracycline Signup and view all the answers

Which of the following features is NOT required to facilitate cloning into a vector?

Multiple cloning sites Signup and view all the answers

Transformation is the process of introducing foreign DNA into a host bacterium.

True Signup and view all the answers

How does increasing the copy number of a vector benefit cloning procedures?

It allows for the recovery of more copies of the target DNA. Signup and view all the answers

Match the following cloning vector components with their functions:

Origin of replication (ori) = Initiates DNA replication Selectable marker = Identifies transformants Cloning sites = Links alien DNA Restriction enzymes = Cuts DNA at specific sequences Signup and view all the answers

What innovation did Herbert Boyer discover regarding DNA strands?

He identified that restriction enzymes leave sticky ends. Signup and view all the answers

Biotechnology is only applicable to modern genetically modified organisms.

False Signup and view all the answers

What was the primary breakthrough that Boyer and Cohen achieved together?

They combined DNA splicing with plasmid technology to create recombinant DNA. Signup and view all the answers

Boyer was born in _____ Pennsylvania.

western Signup and view all the answers

Match the following scientists with their contributions to biotechnology:

Herbert Boyer = Restriction enzymes and sticky ends Stanley Cohen = Plasmid technology E. coli = Bacterial DNA source Biotechnology = Using organisms for production Signup and view all the answers

What is one of the main applications of biotechnology as defined in the content?

Producing useful products using live organisms Signup and view all the answers

Restriction enzymes are capable of cutting DNA at any random location.

False Signup and view all the answers

In what year did Herbert Boyer complete his graduate studies?

1963 Signup and view all the answers

What do restriction endonucleases primarily cut?

The sugar-phosphate backbone of DNA Signup and view all the answers

Sticky ends are produced when restriction enzymes cut DNA at the center of the palindrome sites.

False Signup and view all the answers

Define a palindromic sequence in DNA.

A sequence of base pairs that reads the same on both strands in the same orientation. Signup and view all the answers

Restriction endonucleases form __________ molecules of DNA composed of DNA from different sources.

recombinant Signup and view all the answers

Match the restriction enzyme with its recognition sequence:

EcoRI = GAATTC HindIII = AAGCTT BamHI = GGATCC XhoI = CTCGAG Signup and view all the answers

What is the significance of sticky ends formed after a restriction enzyme cuts DNA?

They facilitate the binding of complementary DNA fragments. Signup and view all the answers

Restriction enzymes can cut DNA at multiple different sites on the same molecule.

True Signup and view all the answers

What enzyme is used to join DNA fragments together after they have been cut with restriction enzymes?

DNA ligase Signup and view all the answers

What enzyme is responsible for joining the ends of cut DNA molecules in recombinant DNA technology?

DNA ligase Signup and view all the answers

An alien piece of DNA can replicate without being part of a chromosome.

False Signup and view all the answers

What is the term for making multiple identical copies of a DNA template?

cloning Signup and view all the answers

The first recombinant DNA was constructed by linking a gene encoding antibiotic resistance with a native __________ of Salmonella typhimurium.

plasmid Signup and view all the answers

Match the following components with their functions in recombinant DNA technology:

Restriction enzymes = Cutting DNA at specific locations Plasmid = Vector for DNA transfer DNA ligase = Joining cut DNA fragments E. coli = Host for cloning antibiotic resistance gene Signup and view all the answers

What is referred to as the origin of replication in a chromosome?

A specific DNA sequence that begins DNA replication Signup and view all the answers

Cloning of the antibiotic resistance gene in E. coli was achieved without the need for a vector.

False Signup and view all the answers

Who were the two scientists that accomplished the construction of the first recombinant DNA?

Stanley Cohen and Herbert Boyer Signup and view all the answers

Which of the following is the first basic step in genetically modifying an organism?

Identification of DNA with desirable genes Signup and view all the answers

Restriction enzymes only function at specific sites within the DNA.

True Signup and view all the answers

What is the function of ligases in recombinant DNA technology?

To join DNA fragments together Signup and view all the answers

The enzyme EcoRI is derived from the bacteria __________.

Escherichia coli Signup and view all the answers

Match the following restriction enzymes with their recognition sequences:

Hind II = Six base pairs EcoRI = GAATTC BamHI = GGATCC NotI = GCGGCCGC Signup and view all the answers

Which of the following statements about exonucleases is true?

They can only remove nucleotides from the ends of the DNA. Signup and view all the answers

Restriction enzymes are a type of exonuclease.

False Signup and view all the answers

Name a common application of recombinant DNA technology.

Producing insulin Signup and view all the answers

What is the primary purpose of a bioreactor?

To provide optimal growth conditions for product production Signup and view all the answers

A sparged stirred-tank bioreactor uses only stirring to mix its contents.

False Signup and view all the answers

What processes are referred to as downstream processing?

Separation and purification of the product Signup and view all the answers

A stirred-tank reactor is usually __________ or with a curved base to facilitate mixing.

cylindrical Signup and view all the answers

Match the components of a bioreactor with their functions:

Agitator system = Facilitates mixing Oxygen delivery system = Supplies oxygen for growth Foam control system = Prevents excessive foam formation Temperature control system = Maintains the desired temperature Signup and view all the answers

Which term best describes the combination of processes like separation, purification, and quality control testing?

Downstream processing Signup and view all the answers

Quality control testing is the same for all biotechnological products.

False Signup and view all the answers

Which method uses high velocity micro-particles coated with DNA to transfer genetic material into plant cells?

Biolistics Signup and view all the answers

Recombinant DNA technology allows for the alteration of __________ to produce genetically modified organisms.

DNA Signup and view all the answers

Isolation of DNA requires breaking open the cell to free it from other macromolecules.

True Signup and view all the answers

What is the role of restriction endonucleases in recombinant DNA technology?

To fragment DNA at specific sequences. Signup and view all the answers

After treating with ________, RNA can be removed from the DNA preparation.

ribonuclease Signup and view all the answers

Match the following enzymes with their specific functions:

Lysozyme = Breaks down bacterial cell walls Cellulase = Degrades cellulose in plant cells Chitinase = Targets fungal cell walls Protease = Breaks down proteins Signup and view all the answers

Which step comes first in the processes of recombinant DNA technology?

Isolation of DNA Signup and view all the answers

The process of culturing host cells ensures the replication of recombinant DNA.

True Signup and view all the answers

What is the purpose of adding chilled ethanol during DNA precipitation?

To precipitate the purified DNA. Signup and view all the answers

What is the primary function of DNA ligase in recombinant DNA technology?

To join together DNA fragments Signup and view all the answers

Eukaryotic cells contain restriction endonucleases.

False Signup and view all the answers

What is the first step in the process of genetically modifying an organism?

Identification of DNA with desirable genes Signup and view all the answers

What are bioreactors primarily used for in large-scale production?

To grow microorganisms or cells under controlled conditions. Signup and view all the answers

Restriction enzymes only cut DNA at specific sequences.

True Signup and view all the answers

The final step in the recombinant DNA process is making a suitable formulation for __________.

marketing Signup and view all the answers

Match the following restriction enzymes with their characteristics:

Hind II = Recognizes a specific sequence of six base pairs EcoRI = Isolated from E. coli Bal I = Creates sticky ends Bam HI = Cuts DNA at a specific recognition site Signup and view all the answers

What type of nucleases remove nucleotides from the ends of DNA?

Exonucleases Signup and view all the answers

All restriction enzymes work on the same recognition sequences.

False Signup and view all the answers

What is the primary role of vectors in recombinant DNA technology?

To carry foreign DNA into host cells. Signup and view all the answers

Which of the following is a requirement for a good cloning vector?

Selectable marker Signup and view all the answers

Plasmids usually contain high copy numbers of their genome within bacterial cells.

False Signup and view all the answers

What is the purpose of the origin of replication (ori) in a cloning vector?

To initiate the replication of linked DNA within host cells. Signup and view all the answers

Selectable markers in cloning vectors are typically resistance genes to antibiotics such as ampicillin and __________.

kanamycin Signup and view all the answers

Match the following vector features with their functions:

Origin of replication = Controls copy number Selectable marker = Identifies transformants Cloning sites = Allows linking of foreign DNA High copy number vector = Increases DNA yield Signup and view all the answers

What type of DNA segments does the cloning vector need to have to link foreign DNA effectively?

Few, preferably single, recognition sites for enzymes Signup and view all the answers

Bacteriophages typically replicate within bacterial cells under the control of chromosomal DNA.

False Signup and view all the answers

Briefly explain what transformation is in the context of cloning vectors.

Transformation is the procedure of introducing a piece of foreign DNA into a host bacterium. Signup and view all the answers

What molecule is used to increase the efficiency of DNA uptake in bacterial cells?

Calcium Signup and view all the answers

Agrobacterium tumefaciens can be used to transfer genes to both plant and animal cells.

False Signup and view all the answers

What is the purpose of heat shock in making bacterial cells competent?

To facilitate the uptake of recombinant DNA. Signup and view all the answers

The _____ plasmid of Agrobacterium tumefaciens is modified to create a cloning vector.

Ti Signup and view all the answers

Match the following methods with their functions:

Heat Shock = Increases DNA uptake in cells Calcium Treatment = Makes bacterial cells competent Agrobacterium tumefaciens = Transfers genes to plants Retroviruses = Transfers genes to animal cells Signup and view all the answers

What happens to colonies that do not produce any color due to insertional inactivation?

They are identified as recombinant colonies. Signup and view all the answers

Name one method, other than heat shock, that can be used to introduce DNA into host cells.

Electroporation or microinjection. Signup and view all the answers

DNA can freely pass through the hydrophobic cell membrane of bacteria.

False Signup and view all the answers

Which method involves directly injecting recombinant DNA into the nucleus of an animal cell?

Micro-injection Signup and view all the answers

The process of culturing host cells occurs before the transfer of recombinant DNA into the host.

False Signup and view all the answers

What is the purpose of using lysozyme when isolating DNA from bacterial cells?

To break down the bacterial cell wall. Signup and view all the answers

Micro-particles of __________ coated with DNA are used in the biolistics method.

gold or tungsten Signup and view all the answers

Match the following steps in recombinant DNA technology with their descriptions:

Isolation of DNA = Obtaining pure DNA from cells Fragmentation = Cutting DNA into smaller pieces using enzymes Ligation = Joining DNA fragments with vectors Transformation = Introducing recombinant DNA into host cells Signup and view all the answers

Which of the following enzymes is used to remove proteins during DNA isolation?

Protease Signup and view all the answers

What is the main requirement for DNA to be cut by restriction enzymes?

It must be in pure form, free from other macromolecules. Signup and view all the answers

What is the primary function of a stirred-tank bioreactor?

To facilitate even mixing and oxygen availability Signup and view all the answers

Extraction of the desired product occurs before culturing the host cells during recombinant DNA technology.

False Signup and view all the answers

Downstream processing includes only the purification of the product.

False Signup and view all the answers

What is one key component of a bioreactor that ensures optimal growth conditions?

Temperature control system Signup and view all the answers

A __________ control system in a bioreactor helps maintain the acidity or alkalinity of the culture.

pH Signup and view all the answers

Match the following components of a bioreactor with their functions:

Agitator system = Ensures even mixing Oxygen delivery system = Provides oxygen to the culture Foam control system = Reduces foam formation Sampling ports = Allows withdrawal of culture samples Signup and view all the answers

Which statement best describes downstream processing?

It includes processes such as purification and quality testing. Signup and view all the answers

A sparged stirred-tank bioreactor does not require any oxygen delivery system.

False Signup and view all the answers

What must products undergo to ensure safety and effectiveness before marketing?

Clinical trials Signup and view all the answers

What is the main role of agarose gel electrophoresis in DNA manipulation?

To visualize DNA fragments Signup and view all the answers

The enzyme DNA polymerase is used during the amplification of DNA in PCR.

True Signup and view all the answers

What is the outcome of adding ligase to the mixture of cut source DNA and cut vector DNA?

Recombinant DNA Signup and view all the answers

PCR can amplify a segment of DNA to approximately ______ copies.

1 billion Signup and view all the answers

Match the following stages of the polymerase chain reaction (PCR) with their functions:

Denaturation = Separating the DNA strands Primer annealing = Binding of primers to the template DNA Extension of primers = Synthesis of new DNA strands Signup and view all the answers

Which type of DNA is primarily used as a template in PCR?

Genomic DNA Signup and view all the answers

Recipient cells must be made 'competent' to take up surrounding DNA.

True Signup and view all the answers

The thermostable DNA polymerase used in PCR is isolated from the bacterium ______.

Thermus aquaticus Signup and view all the answers

What is the primary purpose of using a selectable marker in recombinant DNA technology?

To allow for selection of transformed cells Signup and view all the answers

The expression of foreign genes in host cells is a straightforward process without any technical details.

False Signup and view all the answers

What is the role of bioreactors in biotechnology?

Bioreactors provide a controlled environment for growing large volumes of cultures to produce desired proteins. Signup and view all the answers

What is a palindrome in the context of DNA?

A sequence of base pairs that reads the same on both strands Signup and view all the answers

The ultimate aim of recombinant technologies is to produce a desirable __________.

protein Signup and view all the answers

Restriction endonucleases recognize only non-palindromic sequences in DNA.

False Signup and view all the answers

What are the overhanging stretches formed after restriction enzymes cut the DNA called?

sticky ends Signup and view all the answers

Restriction enzymes cut DNA at specific points in their __________ backbones.

sugar-phosphate Signup and view all the answers

Which of the following is a key advantage of large-scale culturing methods?

It maximizes yield of the desired protein. Signup and view all the answers

Transformed cells will die when spread on agar plates containing ampicillin.

False Signup and view all the answers

What is the role of DNA ligase in recombinant DNA technology?

To bind DNA fragments end-to-end Signup and view all the answers

Why is it necessary to optimize conditions for inducing the expression of target proteins?

To ensure efficient production and proper folding of the desired protein. Signup and view all the answers

Restriction endonucleases must cut the vector and source DNA with the same enzyme to create recombinant DNA.

True Signup and view all the answers

What must happen for recombinant molecules of DNA to be formed?

DNA must be cut by the same restriction enzyme. Signup and view all the answers

What is the purpose of a stirrer in a stirred-tank bioreactor?

To facilitate even mixing and oxygen availability Signup and view all the answers

What type of bioreactor is commonly used to ensure optimal growth conditions?

Stirred-tank bioreactor Signup and view all the answers

A bioreactor provides optimum growth conditions such as temperature, pH, and __________.

substrate Signup and view all the answers

Which of the following processes is part of downstream processing?

Separation and purification Signup and view all the answers

What is the primary purpose of having a selectable marker in a cloning vector?

To identify and eliminate non-transformants Signup and view all the answers

Quality control testing is the same for every biotechnology product.

False Signup and view all the answers

What is recombinant DNA technology commonly used for?

Genetic engineering Signup and view all the answers

Plasmids can have a variable copy number per cell, ranging from one to hundreds.

True Signup and view all the answers

Name one commonly used selectable marker in E. coli.

Ampicillin resistance Signup and view all the answers

The ______ of replication (ori) is responsible for controlling the copy number of linked DNA.

origin Signup and view all the answers

Match the following vector features with their purposes:

Origin of replication (ori) = Controls the replication of the vector Selectable marker = Identifies successful transformants Cloning sites = Facilitates insertion of foreign DNA Signup and view all the answers

Which feature in cloning vectors helps to minimize complications during gene cloning?

Few recognition sites for restriction enzymes Signup and view all the answers

Bacteriophages typically have a lower copy number of their genome compared to plasmids.

False Signup and view all the answers

What is the role of restriction enzymes in the context of cloning vectors?

To cut DNA at specific recognition sites Signup and view all the answers

What is the role of agarose in gel electrophoresis?

It acts as a natural polymer for separating DNA fragments based on size. Signup and view all the answers

DNA fragments can be visualized without any staining technique.

False Signup and view all the answers

What is the common visualization technique used for DNA fragments in gel electrophoresis?

Ethidium bromide staining Signup and view all the answers

What is the result of exposing ethidium bromide stained gel to UV light?

Bright orange colored bands of DNA become visible. Signup and view all the answers

Smaller DNA fragments move farther than larger fragments during gel electrophoresis.

True Signup and view all the answers

What is the procedure called to extract DNA from the agarose gel after electrophoresis?

Elution Signup and view all the answers

What is one of the primary advantages of using bioreactors in large-scale production?

Better control of environmental conditions Signup and view all the answers

Eukaryotic cells do not possess restriction endonucleases.

False Signup and view all the answers

Name one function of a reporter enzyme in genetic engineering.

To monitor the transformation of host cells by foreign DNA. Signup and view all the answers

Restriction enzymes cut DNA at specific __________ sequences.

recognition Signup and view all the answers

What aspect of biotechnology does genetic engineering specifically focus on?

Altering the chemistry of genetic material Signup and view all the answers

Bioprocess engineering ensures the growth of both desired and undesired microbes in biotechnological processes.

False Signup and view all the answers

What is the main advantage of genetic engineering over traditional hybridization methods?

It allows for the isolation and introduction of only desirable genes. Signup and view all the answers

The definition of biotechnology by the European Federation of Biotechnology (EFB) includes the integration of natural science and __________.

organisms Signup and view all the answers

Which of the following is a core technique of biotechnology that maintains a sterile environment?

Bioprocess engineering Signup and view all the answers

Match each biotechnology term with its correct description:

Genetic engineering = Techniques to alter genetic material Bioprocess engineering = Ensures sterile conditions in production Recombinant DNA = DNA formed from two different sources Gene cloning = Copying a specific gene for study or use Signup and view all the answers

A piece of DNA transferred into an alien organism is guaranteed to replicate in the progeny cells.

False Signup and view all the answers

What is one significant benefit of sexual reproduction compared to asexual reproduction in terms of genetic variation?

It provides opportunities for variations and unique genetic combinations. Signup and view all the answers

Which method is used to directly inject recombinant DNA into the nucleus of an animal cell?

Micro-injection Signup and view all the answers

Isolation of DNA involves breaking open the cell to release DNA and other macromolecules.

True Signup and view all the answers

What type of enzymes are used to remove RNA during DNA isolation?

Ribonuclease Signup and view all the answers

The method known as __________ involves bombarding plant cells with DNA-coated micro-particles.

biolistics Signup and view all the answers

Chilled ethanol is used to precipitate purified DNA after isolation.

True Signup and view all the answers

Name one type of enzyme used to break down bacterial cell walls during DNA isolation.

Lysozyme Signup and view all the answers

What is the main role of the selectable marker in a cloning vector?

To eliminate non-transformants and permit growth of transformants Signup and view all the answers

Bacteriophages typically have a lower copy number of their genome within bacterial cells than plasmids.

False Signup and view all the answers

What is the function of the origin of replication (ori) in a cloning vector?

It initiates replication of linked DNA within host cells. Signup and view all the answers

The presence of single recognition sites for restriction enzymes in a vector minimizes the generation of __________ during cloning.

fragments Signup and view all the answers

What is the primary function of a restriction enzyme during DNA manipulation?

To cut DNA at specific locations Signup and view all the answers

Agarose gel electrophoresis is used to check the progression of a restriction enzyme digestion.

True Signup and view all the answers

Which of the following is a characteristic feature of cloning vectors?

Includes a high copy number origin of replication Signup and view all the answers

What is the purpose of using ligase in the process of recombinant DNA preparation?

To join the DNA fragments together Signup and view all the answers

Match the following features of cloning vectors with their descriptions:

Origin of replication = Controls the replication of linked DNA Selectable marker = Helps in identifying transformants Cloning sites = Allows integration of alien DNA Signup and view all the answers

Transformation refers to the procedure of successfully isolating recombinant DNA.

False Signup and view all the answers

The high-temperature step in PCR is known as _____, which separates the double-stranded DNA.

denaturation Signup and view all the answers

Match the following steps of the Polymerase Chain Reaction (PCR) with their correct descriptions:

Denaturation = Separating the DNA strands Primer annealing = Binding of primers to the DNA Extension = Synthesis of new DNA strands Thermal cycling = Repeating the cycles of denaturation, annealing, and extension Signup and view all the answers

Common selectable markers used for E. coli include genes that provide resistance to __________.

antibiotics Signup and view all the answers

Which of the following enzymes is key for extending the primers during PCR?

DNA polymerase Signup and view all the answers

A recombinant DNA molecule can only be inserted into host cells through one method.

False Signup and view all the answers

What is the term used to describe recipient cells that have been treated to take up DNA?

Competent cells Signup and view all the answers

What method is used to make bacterial cells competent for DNA uptake?

Heat shock and ice incubation Signup and view all the answers

Agrobacterium tumefaciens is used to deliver genes into animal cells.

False Signup and view all the answers

What DNA component does Agrobacterium tumefaciens deliver to transform plant cells?

T-DNA Signup and view all the answers

The process of introducing foreign DNA into host cells is called ______.

transformation Signup and view all the answers

Match the following vectors with their uses:

Agrobacterium tumefaciens = Gene delivery in plants Retroviruses = Gene delivery in animals Plasmids = Cloning in bacteria Bacteriophages = Gene delivery in bacterial cells Signup and view all the answers

Which of the following statements is true regarding recombinant colonies?

They do not produce color due to insertional inactivation. Signup and view all the answers

The rationale behind using pathogens for gene delivery is based on their natural abilities to transform host cells.

True Signup and view all the answers

To facilitate the entry of DNA, bacterial cells are treated with a specific concentration of __________.

divalent cation Signup and view all the answers

What was the main focus of Herbert Boyer's studies in 1969?

Restriction enzymes of E.coli Signup and view all the answers

Biotechnology can be broadly defined as the use of natural organisms for various applications.

True Signup and view all the answers

What are the small ringlets of DNA called that are used in gene cloning?

plasmids Signup and view all the answers

The process of recombining segments of DNA is known as __________.

DNA splicing Signup and view all the answers

Match each scientist with their contribution to biotechnology:

Herbert Boyer = Studied restriction enzymes Stanley Cohen = Developed plasmid insertion methods Restriction enzymes = Cutting DNA at specific sequences Biotechnology = Application of organisms for products Signup and view all the answers

What is an essential feature introduced by Boyer and Cohen in their biotechnology work?

Recombination of DNA Signup and view all the answers

Genetically modified organisms are a key component of modern biotechnology.

True Signup and view all the answers

In what year did Herbert Boyer complete his graduate work at the University of Pittsburgh?

1963 Signup and view all the answers

What is the primary purpose of using a restriction enzyme in DNA manipulation?

To cut DNA at specific locations Signup and view all the answers

Agarose gel electrophoresis can be used to confirm the successful cutting of DNA by restriction enzymes.

True Signup and view all the answers

What type of enzyme is used to ligate cut DNA segments together during recombinant DNA preparation?

Ligase Signup and view all the answers

The Polymerase Chain Reaction (PCR) uses a thermostable DNA polymerase from the organism __________ to extend primers.

Thermus aquaticus Signup and view all the answers

Match the following PCR steps with their correct descriptions:

Denaturation = Separation of double-stranded DNA into single strands Primer annealing = Attachment of primers to the DNA template Extension of primers = Synthesis of new DNA strands from primers Signup and view all the answers

What is created after the amplification of a gene of interest using PCR?

Billion copies of the target gene Signup and view all the answers

Recipient cells must be made 'competent' before they can take up recombinant DNA.

True Signup and view all the answers

What type of bond does DNA ligase form when joining DNA fragments?

Phosphodiester bond Signup and view all the answers

What is the purpose of using restriction endonucleases in recombinant DNA technology?

To cut DNA at specific sequences Signup and view all the answers

Bioreactors are primarily used for the production of therapeutic proteins on a small scale.

False Signup and view all the answers

Name one advantage of using stirred tank bioreactors over shake flasks.

Better aeration properties Signup and view all the answers

The functional protein produced after isolating foreign DNA is called the __________.

gene product Signup and view all the answers

Match each component of recombinant DNA technology with its function:

Restriction endonucleases = Isolate DNA fragments DNA ligase = Join DNA fragments together Plasmid vectors = Transport foreign DNA into host cells Expression systems = Produce gene products Signup and view all the answers

Heterologous expression refers to expressing a protein in the same organism where the gene was cloned.

False Signup and view all the answers

What are bioreactors used for?

To process large volumes of culture to produce specific products. Signup and view all the answers

The cells can be maintained in their physiologically most active _____ phase for optimal growth.

log/exponential Signup and view all the answers

Why is large-scale production of recombinant proteins necessary?

To ensure that sufficient quantities are available for research or therapeutic use Signup and view all the answers

Ampicillin-resistant cells will die when exposed to ampicillin.

False Signup and view all the answers

What happens to foreign DNA when it is inserted into a cloning vector and transferred into host cells?

It gets multiplied. Signup and view all the answers

What is the primary purpose of gel electrophoresis in DNA analysis?

To separate DNA fragments by size Signup and view all the answers

DNA fragments can be seen with the naked eye without any staining technique.

False Signup and view all the answers

What is the role of ethidium bromide in gel electrophoresis?

It stains DNA for visualization under UV light. Signup and view all the answers

The agarose gel provides a __________________ effect that allows DNA fragments to separate based on size.

sieving Signup and view all the answers

Match the following terms with their descriptions related to gel electrophoresis:

Anode = Positive electrode where DNA fragments migrate towards Agarose = Matrix used in gel electrophoresis Ethidium bromide = Staining agent for DNA visualization Elution = Process of extracting DNA from the gel Signup and view all the answers

How does the size of DNA fragments affect their movement in gel electrophoresis?

Smaller fragments move farther than larger ones Signup and view all the answers

Elution is the process of cutting DNA bands from the agarose gel.

True Signup and view all the answers

What is a primary aspect of genetic engineering?

Altering the chemistry of genetic material Signup and view all the answers

Bioprocess engineering focuses on maintaining a sterile environment for the growth of desired microbes or cells.

True Signup and view all the answers

Define biotechnology based on the EFB's definition.

The integration of natural science and organisms for products and services. Signup and view all the answers

Traditional hybridization processes often lead to the inclusion of __________ genes.

undesirable Signup and view all the answers

Match the following biotechnology techniques with their descriptions:

Genetic engineering = Techniques to alter genetic material Bioprocess engineering = Maintaining a sterile environment Recombinant DNA = Combination of DNA from different sources Gene cloning = Making copies of a specific gene Signup and view all the answers

What is one of the core benefits of sexual reproduction over asexual reproduction?

Opportunities for genetic variation Signup and view all the answers

Only a piece of DNA that is integrated into an alien organism can guarantee its multiplication in progeny cells.

False Signup and view all the answers

What two techniques form the foundation of modern biotechnology?

Genetic engineering and bioprocess engineering. Signup and view all the answers

What is the role of DNA ligase in the process of genetic engineering?

To join DNA fragments together Signup and view all the answers

Eukaryotic cells possess restriction endonucleases.

False Signup and view all the answers

What is the main advantage of using bioreactors for large scale production?

Better aeration and mixing properties Signup and view all the answers

A ___________ is used to monitor the transformation of host cells by foreign DNA.

reporter enzyme Signup and view all the answers

Match the following components of genetic engineering with their functions:

Restriction endonucleases = Cutting DNA at specific sequences DNA ligase = Joining DNA fragments Bioreactors = Supporting large-scale cell growth Selectable markers = Identifying successful transformations Signup and view all the answers

Which method involves the direct injection of recombinant DNA into an animal cell?

Micro-injection Signup and view all the answers

The first step in recombinant DNA technology is the transfer of recombinant DNA into the host.

False Signup and view all the answers

What enzyme is specifically used to remove RNA during the isolation of DNA?

Ribonuclease Signup and view all the answers

The process of breaking open cells to release DNA is called ______.

cell lysis Signup and view all the answers

Match the following methods with their descriptions:

Micro-injection = Directly injects DNA into the nucleus of animal cells Biolistics = Uses high-velocity particles coated with DNA Pathogen vectors = Infects the host cell to introduce DNA Signup and view all the answers

Which molecule is NOT typically removed during DNA isolation?

Nucleotides Signup and view all the answers

Ethanol is used to precipitate purified DNA during isolation.

True Signup and view all the answers

What are the two main types of molecules that must be removed from DNA during its isolation?

Proteins and RNA Signup and view all the answers

What is the primary purpose of gel electrophoresis?

To separate DNA fragments based on size Signup and view all the answers

What compound is commonly used to stain DNA for visualization in gel electrophoresis?

ethidium bromide Signup and view all the answers

The larger the DNA fragment, the ______ it moves through the agarose gel during electrophoresis.

less Signup and view all the answers

Match the following gel electrophoresis components with their functions:

Agarose = Matrix for separating DNA Electric field = Facilitates DNA movement Ethidium bromide = Stains DNA for visualization UV light = Enables visualization of stained DNA Signup and view all the answers

What happens to DNA fragments after gel electrophoresis?

They are cut out and extracted from the gel Signup and view all the answers

What is the process of extracting DNA bands from agarose gel called?

elution Signup and view all the answers

What is the primary role of restriction enzymes in genetic engineering?

To cut DNA strands at specific sequences Signup and view all the answers

Herbert Boyer was primarily focused on studying plant DNA.

False Signup and view all the answers

What discovery allowed Boyer and Cohen to recombine segments of DNA?

The use of restriction enzymes to create sticky ends. Signup and view all the answers

Biotechnology deals with techniques of using live organisms or ________ from organisms to produce useful products.

enzymes Signup and view all the answers

Match the following individuals to their contributions in biotechnology:

Herbert Boyer = Co-developed recombinant DNA technology Stanley Cohen = Developed plasmid reinsertion method Restriction Enzymes = Cut DNA at specific sites Plasmids = Small ringlets of DNA in bacteria Signup and view all the answers

Which of the following represents a traditional application of biotechnology?

Making wine Signup and view all the answers

Plasmids can replicate independently of the bacterial genome.

True Signup and view all the answers

What is the foundational significance of the work done by Boyer and Cohen?

It laid the groundwork for the field of biotechnology. Signup and view all the answers

Which feature is crucial for controlling the copy number of linked DNA in a cloning vector?

Origin of replication (ori) Signup and view all the answers

Bacteriophages typically have a lower copy number of their genome within bacterial cells compared to plasmids.

False Signup and view all the answers

What is the purpose of a selectable marker in a cloning vector?

To identify and eliminate non-transformants and allow for the growth of transformants. Signup and view all the answers

The genes encoding resistance to antibiotics such as _________ are commonly used as selectable markers in E. coli.

ampicillin Signup and view all the answers

Match the following cloning vector features to their functions:

Origin of replication = Controls the copy number Selectable marker = Identifies transformants Cloning site = Links foreign DNA Restriction enzyme site = Facilitates cutting of DNA Signup and view all the answers

Which of the following describes the role of cloning sites in a vector?

They should have few recognition sites for restriction enzymes. Signup and view all the answers

Plasmids are known to replicate within bacterial cells independently of chromosomal DNA.

True Signup and view all the answers

What is transformation in the context of cloning vectors?

The process of introducing a piece of DNA into a host bacterium. Signup and view all the answers

Which technique involves altering the chemistry of genetic material to change an organism's phenotype?

Genetic Engineering Signup and view all the answers

Bioprocess engineering focuses on creating a sterile environment to promote the growth of any microbial organism.

False Signup and view all the answers

The definition of biotechnology given by the European Federation of Biotechnology is the integration of natural science and organisms, cells, parts thereof, and molecular __________ for products and services.

analogues Signup and view all the answers

Match the following concepts with their descriptions:

Genetic Engineering = Techniques to alter DNA and RNA Bioprocess Engineering = Maintenance of sterile conditions for microbial growth Recombinant DNA = DNA that has been artificially made using genetic engineering Selectable Marker = A gene that allows for the identification of cells that have taken up foreign DNA Signup and view all the answers

What is the likely outcome of transferring a piece of DNA into an alien organism?

It will likely not be able to replicate in progeny cells. Signup and view all the answers

Sexual reproduction typically diminishes genetic variation in organisms.

False Signup and view all the answers

In biotechnology, the term _________ refers to the process of transferring foreign DNA into a host organism.

transformation Signup and view all the answers

The isolation of genetic material requires that DNA be free from other macromolecules.

True Signup and view all the answers

The process of __________ involves treating cells with enzymes to release DNA.

cell lysis Signup and view all the answers

Match the following DNA processing steps with their descriptions:

Isolation of DNA = Release DNA from cells Fragmentation of DNA = Cutting DNA into smaller pieces Ligation = Joining DNA fragments together Transformation = Inserting DNA into a host organism Signup and view all the answers

What type of cells can be treated with lysozyme for DNA isolation?

Bacterial cells Signup and view all the answers

Ethanol is used to precipitate DNA during its purification process.

True Signup and view all the answers

Which enzymes can be used to remove RNA during the DNA isolation process?

Ribonucleases Signup and view all the answers

What enzyme is primarily responsible for extending primers in the Polymerase Chain Reaction (PCR)?

DNA polymerase Signup and view all the answers

Agarose gel electrophoresis can be used to check the progression of a restriction enzyme digestion.

True Signup and view all the answers

What is the term for the mixed DNA and vector after ligation in recombinant DNA technology?

recombinant DNA Signup and view all the answers

The __________ temperature polymerase is used in PCR to remain active during high temperature denaturation.

thermostable Signup and view all the answers

Match the following steps of PCR with their corresponding functions:

Denaturation = Separation of double-stranded DNA Primer annealing = Binding of primers to the template DNA Extension of primers = Synthesis of new DNA strands Cycle repetition = Amplification of DNA segments Signup and view all the answers

Which of the following best describes what happens during the primer annealing step in PCR?

Primers bind to the double-stranded DNA Signup and view all the answers

Competent cells require treatment before they can take up recombinant DNA from their surroundings.

True Signup and view all the answers

What is the main purpose of genetic engineering?

To alter the chemistry of genetic material Signup and view all the answers

Asexual reproduction allows for greater genetic variation compared to sexual reproduction.

False Signup and view all the answers

What is the process called that introduces ligated DNA into host cells?

transformation Signup and view all the answers

Bioprocess engineering ensures the growth of only the desired __________ in large quantities for manufacturing bioproducts.

microbe/eukaryotic cell Signup and view all the answers

Match the following biotechnology terms with their definitions:

Genetic engineering = Altering the chemistry of DNA/RNA Recombinant DNA = DNA that is artificially formed by combining constituents from different organisms Bioprocess engineering = Maintaining sterile conditions for microbial growth Gene transfer = Introducing genes from one organism to another Signup and view all the answers

What enzyme is primarily responsible for joining cut DNA molecules?

DNA ligase Signup and view all the answers

Which organization defined biotechnology in a way that includes both traditional and modern methods?

European Federation of Biotechnology Signup and view all the answers

If a piece of DNA is transferred into an alien organism, it will always multiply in the progeny cells.

False Signup and view all the answers

The origin of replication in DNA is essential for initiating replication.

True Signup and view all the answers

Who were the two scientists credited with constructing the first recombinant DNA molecule?

Stanley Cohen and Herbert Boyer Signup and view all the answers

What are two core techniques that enabled the birth of modern biotechnology?

Genetic engineering and bioprocess engineering. Signup and view all the answers

Recombinant DNA is created when an alien piece of DNA is integrated into ______.

a plasmid Signup and view all the answers

Match the following DNA components with their functions:

Restriction Enzyme = Cuts DNA at specific locations Plasmid = Vector for DNA insertion DNA Ligase = Joins DNA fragments Origin of Replication = Initiates DNA replication Signup and view all the answers

What medium is most commonly used for gel electrophoresis?

Agarose Signup and view all the answers

What is the primary application of inserting an antibiotic resistance gene into E. coli?

To enable cloning of the gene Signup and view all the answers

DNA fragments can be visualized under visible light without any staining.

False Signup and view all the answers

When integrated into the host genome, an alien piece of DNA cannot be inherited.

False Signup and view all the answers

What type of DNA is also known as autonomously replicating circular extra-chromosomal DNA?

Plasmid Signup and view all the answers

What process is used to separate DNA fragments in gel electrophoresis?

Gel electrophoresis Signup and view all the answers

The smaller the DNA fragment size, the farther it moves through the __________ gel.

agarose Signup and view all the answers

Which compound is used to stain DNA for visualization after gel electrophoresis?

Ethidium bromide Signup and view all the answers

What is the process called when separated DNA bands are cut from the agarose gel?

Elution Signup and view all the answers

DNA fragments stain bright green when exposed to UV light after using ethidium bromide.

False Signup and view all the answers

What is the second step in genetically modifying an organism?

Introduction of identified DNA into the host Signup and view all the answers

Restriction endonucleases can only cut DNA at the ends of the molecules.

False Signup and view all the answers

What term describes the specific base sequence that restriction enzymes recognize?

Recognition sequence Signup and view all the answers

EcoRI is a restriction enzyme derived from __________.

Escherichia coli Signup and view all the answers

Which enzyme is primarily responsible for making cuts within the DNA?

Restriction endonucleases Signup and view all the answers

All restriction enzymes recognize the same nucleotide sequences.

False Signup and view all the answers

What type of nucleases do endonucleases belong to?

Nucleases Signup and view all the answers

What is the result of ligating foreign DNA at the BamH I site of the tetracycline resistance gene?

The gene is inactivated Signup and view all the answers

Non-recombinants can grow on media containing both ampicillin and tetracycline.

True Signup and view all the answers

What is the role of the antibiotic resistance gene in cloning vectors like pBR322?

To select for transformants Signup and view all the answers

The process of insertional inactivation occurs when a foreign DNA is inserted into the gene coding for __________.

β-galactosidase Signup and view all the answers

What color will colonies appear if the plasmid does not have an insert when using a chromogenic substrate?

Blue Signup and view all the answers

Match the following components of cloning vectors with their functions:

ori = Allows plasmid replication ampR = Confers resistance to ampicillin tetR = Confers resistance to tetracycline rop = Involved in plasmid replication control Signup and view all the answers

What is the significance of using alternative selectable markers in recombinant DNA technology?

To simplify the selection of recombinants Signup and view all the answers

The inactivation of the tetracycline resistance gene just involves removing a part of the gene.

False Signup and view all the answers

What is the main purpose of using restriction enzymes in DNA manipulation?

To cut DNA at specific locations Signup and view all the answers

Agarose gel electrophoresis uses a negatively charged DNA molecule to migrate towards the negative electrode.

False Signup and view all the answers

What is the purpose of Polymerase Chain Reaction (PCR)?

To amplify a specific segment of DNA. Signup and view all the answers

A ______ is added to the mixture of the cut DNA and vector to create recombinant DNA.

ligase Signup and view all the answers

Match the following steps of PCR with their descriptions:

Denaturation = Separation of the DNA strands at high temperature Primer annealing = Binding of primers to the template DNA Extension = Synthesis of new DNA strands by DNA polymerase Signup and view all the answers

Which DNA polymerase is used in PCR because of its thermostability?

Taq polymerase Signup and view all the answers

Recipient cells must be made ‘competent’ to take up the introduced DNA.

True Signup and view all the answers

What type of DNA is created when a gene of interest is ligated with a vector?

Recombinant DNA Signup and view all the answers

Which method involves directly injecting recombinant DNA into an animal cell?

Micro-injection Signup and view all the answers

The process of isolating DNA requires breaking open the cell to release it.

True Signup and view all the answers

What enzyme is used to remove RNA during the DNA isolation process?

Ribonuclease Signup and view all the answers

The __________ method uses high-velocity micro-particles coated with DNA for gene transfer.

biolistics Signup and view all the answers

What is the purpose of using chilled ethanol in the DNA isolation process?

To precipitate the DNA Signup and view all the answers

Protease is used to remove lipids from the mixture during DNA isolation.

False Signup and view all the answers

What is the final step of the recombinant DNA technology process mentioned in the content?

Extraction of the desired product Signup and view all the answers

What role do restriction enzymes play in recombinant DNA technology?

They cut DNA at specific locations. Signup and view all the answers

An alien piece of DNA must be integrated into a chromosome to replicate and be inherited.

True Signup and view all the answers

What is the main purpose of DNA ligase in the construction of recombinant DNA?

To join the ends of cut DNA molecules. Signup and view all the answers

The first recombinant DNA was created from a gene encoding __________ resistance.

antibiotic Signup and view all the answers

What is the significance of having an origin of replication in a cloning vector?

It enables the cloning vector to replicate independently. Signup and view all the answers

Transferring recombinant DNA into E. coli can help produce multiple copies of the desired gene.

True Signup and view all the answers

Who were the scientists known for constructing the first recombinant DNA molecule?

Stanley Cohen and Herbert Boyer Signup and view all the answers

Which technique is primarily used to alter the chemistry of genetic material?

Genetic engineering Signup and view all the answers

Bioprocess engineering focuses on maintaining a contamination-free environment in chemical processes.

True Signup and view all the answers

What is the primary purpose of genetic engineering in biotechnology?

To introduce desirable genes into an organism without unwanted genes. Signup and view all the answers

The integration of natural science and organisms for products and services is defined by __________.

biotechnology Signup and view all the answers

Match the following terms to their descriptions:

Genetic engineering = Techniques to alter genetic material Bioprocess engineering = Maintenance of sterile environments Recombinant DNA = DNA from different sources combined Gene cloning = Duplicating specific genes for study Signup and view all the answers

What is one major disadvantage of traditional hybridization methods?

They can include undesirable genes. Signup and view all the answers

A piece of DNA transferred into an alien organism is guaranteed to multiply in its progeny cells.

False Signup and view all the answers

What is the likely fate of a piece of DNA that does not integrate into the host organism's genome?

It will most likely be degraded and not multiply. Signup and view all the answers

What is the purpose of the ampicillin resistance gene in transformed cells?

To allow cells to grow in the presence of ampicillin Signup and view all the answers

A selectable marker can imply that a transformed cell will die in the presence of certain antibiotics.

False Signup and view all the answers

What is produced on a large scale in recombinant DNA technology?

Desirable proteins Signup and view all the answers

In recombinant DNA technology, the foreign gene that is expressed in a host cell is known as a __________ protein.

recombinant Signup and view all the answers

What is the primary benefit of using bioreactors over small volume cultures?

Higher biomass and yields of desired protein Signup and view all the answers

Continuous culture systems can only be used in small-scale production.

False Signup and view all the answers

What type of cell cultures may be used for extracting and purifying a desired protein?

Cloned cells Signup and view all the answers

What significant breakthrough did Boyer and Cohen achieve?

Recombining segments of DNA Signup and view all the answers

Biotechnology is solely focused on genetically modified organisms.

False Signup and view all the answers

What are the 'sticky ends' created by restriction enzymes used for?

They are used for joining together pieces of DNA. Signup and view all the answers

Herbert Boyer was born in the year ____.

1936 Signup and view all the answers

Match the following individuals with their contributions:

Herbert Boyer = Restriction enzymes and DNA splicing Stanley Cohen = Plasmid manipulation E.coli = Source of restriction enzymes Biotechnology = Use of living organisms for useful products Signup and view all the answers

Which process describes the act of reintroducing plasmids into bacterial cells?

Transformation Signup and view all the answers

What method did Cohen develop for handling plasmids?

Removing plasmids from bacterial cells. Signup and view all the answers

Restriction enzymes only cut DNA at specific recognition sites.

True Signup and view all the answers

What is the main function of restriction enzymes in recombinant DNA technology?

To cut DNA at specific sequences Signup and view all the answers

Exonucleases cut DNA at specific internal positions.

False Signup and view all the answers

Name one restriction endonuclease that was characterized in 1968.

Hind II Signup and view all the answers

The process of introducing foreign DNA into a host organism is called __________.

transformation Signup and view all the answers

Match the following types of nucleases with their descriptions:

Exonucleases = Remove nucleotides from the ends of DNA Endonucleases = Cut DNA at specific positions within the strand Signup and view all the answers

Which term describes the sequence that restriction enzymes recognize in DNA?

Recognition sequence Signup and view all the answers

What is the role of Agrobacterium tumefaciens in gene delivery?

It delivers T-DNA to transform plant cells. Signup and view all the answers

The correct naming convention for restriction enzymes includes letters from the name of the genus and species they are isolated from.

True Signup and view all the answers

Retroviruses can be modified to deliver desirable genes into animal cells.

True Signup and view all the answers

What must be done to bacterial cells to make them competent for DNA uptake?

They must be treated with calcium. Signup and view all the answers

The process of introducing recombinant DNA into bacterial cells is called __________.

transformation Signup and view all the answers

Which best describes the Ti plasmid of Agrobacterium tumefaciens?

It is a cloning vector suitable for gene delivery. Signup and view all the answers

DNA cannot pass through cell membranes due to its hydrophilic nature.

True Signup and view all the answers

What is the purpose of making bacterial cells competent?

To enable them to take up foreign DNA. Signup and view all the answers

Bacteriophages typically have a lower copy number of their genome per cell compared to plasmids.

False Signup and view all the answers

What are selectable markers used for in cloning vectors?

To identify and eliminate non-transformants and permit growth of transformants. Signup and view all the answers

A vector requires a selectable marker to help identify and eliminate __________.

non-transformants Signup and view all the answers

Which of the following is a benefit of high copy number vectors?

They allow for the recovery of more copies of target DNA. Signup and view all the answers

Restriction enzymes should ideally have multiple recognition sites to simplify gene cloning.

False Signup and view all the answers

Which antibiotic resistance genes are commonly used as selectable markers in E. coli?

Ampicillin, chloramphenicol, tetracycline, kanamycin Signup and view all the answers

What is the first step in the process of recombinant DNA technology?

Isolation of genetic material Signup and view all the answers

Biolistics uses high-velocity micro-particles to deliver DNA into animal cells.

False Signup and view all the answers

What is the primary purpose of using a restriction enzyme in recombinant DNA technology?

To cut DNA at specific locations Signup and view all the answers

Agarose gel electrophoresis is used to visualize the movement of DNA towards the negative electrode.

False Signup and view all the answers

Name the enzyme that is used to remove RNA during DNA isolation.

Ribonuclease Signup and view all the answers

The method known as __________ involves directly injecting recombinant DNA into the nucleus of an animal cell.

micro-injection Signup and view all the answers

What enzyme is used to join the cut DNA fragments and ligate them together?

Ligase Signup and view all the answers

The technique used to amplify the gene of interest is called _____ .

PCR Signup and view all the answers

Which step follows the isolation of the desired DNA fragment?

Ligation of the DNA fragment into a vector Signup and view all the answers

Match the following steps of the PCR process with their correct descriptions:

Denaturation = Separation of double-stranded DNA into single strands Primer annealing = Binding of primers to the template DNA Extension = Synthesis of new DNA strands by DNA polymerase Signup and view all the answers

Proteins can be removed from the DNA mixture using ribonuclease.

False Signup and view all the answers

Which component is crucial for the functioning of PCR at high temperatures?

Thermostable DNA polymerase Signup and view all the answers

Competent cells are made ready to take up foreign DNA during the transformation process.

True Signup and view all the answers

What is the final result of adding chilled ethanol during DNA isolation?

Purified DNA precipitates out Signup and view all the answers

What is the purpose of adding a gene for antibiotic resistance to a recombinant DNA?

To select successfully transformed cells Signup and view all the answers

What is the role of the origin of replication in a chromosome?

It initiates the replication process. Signup and view all the answers

Plasmids can act as vectors to transfer alien DNA into host organisms.

True Signup and view all the answers

What is recombinant DNA?

A new combination of DNA created by linking foreign DNA with plasmid DNA. Signup and view all the answers

The enzyme __________ is responsible for joining the ends of cut DNA molecules.

DNA ligase Signup and view all the answers

Match the following scientists with their contributions:

Stanley Cohen = Linked antibiotic resistance gene with plasmid Herbert Boyer = Developed the concept of recombinant DNA Restriction Enzymes = Cut DNA at specific sequences DNA Ligase = Joins the ends of cut DNA molecules Signup and view all the answers

In what year did Cohen and Boyer successfully construct recombinant DNA?

1972 Signup and view all the answers

Cutting DNA with restriction enzymes can be done at any location on the DNA strand.

False Signup and view all the answers

Name a common selectable marker used in E. coli.

Antibiotic resistance gene Signup and view all the answers

What is the primary focus of biotechnology today?

Using genetically modified organisms for large scale production Signup and view all the answers

Herbert Boyer discovered the ability of restriction enzymes to cut DNA, leading to the concept of 'sticky ends'.

True Signup and view all the answers

What process combines DNA splicing and the use of plasmids to manipulate genetic material?

Recombination of DNA segments Signup and view all the answers

Herbert Boyer performed studies on restriction enzymes of the __________ bacterium.

E.coli Signup and view all the answers

What does biotechnology broadly encompass?

All forms of product creation by organisms or their enzymes Signup and view all the answers

The primary component of biotechnology is the use of chemical processes in laboratories.

False Signup and view all the answers

What type of DNA structure, invented by Stanley Cohen, is crucial for the replication of genetic material in bacterial cells?

Plasmids Signup and view all the answers

Which enzyme is primarily used to join foreign DNA to vector DNA in recombinant DNA technology?

DNA ligase Signup and view all the answers

Bioreactors are only used for small-scale production of recombinant proteins.

False Signup and view all the answers

What is the role of a restriction enzyme in genetic engineering?

To cut DNA at specific sequences. Signup and view all the answers

The process of producing functional proteins from foreign genes involves the use of ________ such as bioreactors.

bioprocesses Signup and view all the answers

Match the following components of recombinant DNA technology with their functions:

Restriction enzymes = Cut DNA at specific sequences DNA ligase = Join DNA fragments Plasmid vectors = Carry foreign DNA into host cells Bioreactors = Provide suitable conditions for large-scale growth Signup and view all the answers

What is the function of a selectable marker in a cloning vector?

To identify and eliminate non-transformants Signup and view all the answers

Bacteriophages have a higher copy number of their genome within bacterial cells compared to plasmids.

True Signup and view all the answers

Agarose gel electrophoresis is used to check the purity of DNA samples.

False Signup and view all the answers

What is the purpose of ligase in the process of creating recombinant DNA?

To join DNA fragments together Signup and view all the answers

What is the purpose of a selectable marker in recombinant DNA technology?

To select transformed cells in the presence of certain conditions Signup and view all the answers

The genes encoding resistance to antibiotics such as ampicillin and __________ are considered useful selectable markers.

tetracycline Signup and view all the answers

The main goal of recombinant DNA technology is to produce a desirable protein.

True Signup and view all the answers

The enzyme ________ is used to synthesize multiple copies of a gene during PCR.

DNA polymerase Signup and view all the answers

Match the cloning vector components with their functions:

Origin of replication = Controls copy number and replication initiation Selectable marker = Identifies transformants Cloning sites = Facilitates linking foreign DNA Restriction enzyme recognition sites = Allows precise cuts for cloning Signup and view all the answers

Which of the following is a common feature of plasmids used as cloning vectors?

High copy number per bacterial cell Signup and view all the answers

What is produced on a large scale in bioprocess engineering?

Desired protein Signup and view all the answers

Match the following components of PCR to their functions:

Denaturation = Separating double-stranded DNA into single strands Primer annealing = Binding of primers to the template DNA Extension = Synthesis of new DNA strands by DNA polymerase Signup and view all the answers

Bioreactors can be thought of as vessels in which raw materials are biologically converted into specific _______.

products Signup and view all the answers

A vector must have multiple recognition sites for restriction enzymes to aid in gene cloning.

False Signup and view all the answers

What effect does increasing the number of PCR cycles have on the amount of DNA produced?

It amplifies the DNA exponentially Signup and view all the answers

What is the benefit of using plasmids with a high copy number for cloning?

They allow for the production of many copies of the target DNA. Signup and view all the answers

Thermostable DNA polymerase is important in PCR because it can withstand the high temperatures used in denaturation.

True Signup and view all the answers

Describe how recipient cells become 'competent' to receive DNA.

They are treated to allow them to take up DNA from their environment. Signup and view all the answers

What advantage do continuous culture systems provide in protein production?

They allow for consistent removal and addition of culture medium. Signup and view all the answers

Cloning vectors must have at least one selectable marker to function correctly.

True Signup and view all the answers

Name one condition that must be optimized for the expression of a foreign gene.

Temperature Signup and view all the answers

What is the term for the ends created by restriction enzymes after cutting DNA?

Sticky ends Signup and view all the answers

Restriction endonucleases can create blunt ends when they cut DNA.

False Signup and view all the answers

What is a palindrome in the context of DNA sequences?

A sequence of base pairs that reads the same on both strands. Signup and view all the answers

Restriction endonucleases recognize specific __________ sequences in DNA.

palindromic Signup and view all the answers

Match the following restriction enzymes with their actions:

EcoRI = Cuts at GAATTC HindIII = Cuts at AAGCTT BamHI = Cuts at GGATCC NotI = Cuts at GCGGCCGC Signup and view all the answers

What is required for the creation of a recombinant vector molecule?

Cutting both the vector and DNA with the same restriction enzyme Signup and view all the answers

The formation of hydrogen bonds occurs between complementary sticky ends after DNA has been cut.

True Signup and view all the answers

Restriction enzymes can recognize and cut DNA at specific sequences.

True Signup and view all the answers

What does the 'R' in EcoRI signify?

RY 13 Signup and view all the answers

Restriction endonucleases make cuts at specific _______ within the DNA.

positions Signup and view all the answers

Which type of nucleases remove nucleotides from the ends of the DNA?

Exonucleases Signup and view all the answers

All restriction enzymes recognize the same recognition sequence.

False Signup and view all the answers

What is the primary function of restriction endonucleases in genetic engineering?

To cut DNA at specific sequences Signup and view all the answers

Bacteriophages can have higher copy numbers than plasmids within bacterial cells.

True Signup and view all the answers

What is the process of using bioreactors for large scale production in biotechnology?

Bioreactors provide a controlled environment for the growth of microorganisms, allowing for efficient large-scale production of biological products. Signup and view all the answers

In genetic engineering, the purpose of the __________ is to ferry foreign DNA into host organisms.

vector Signup and view all the answers

What does 'ori' stand for in the context of cloning vectors?

Origin of replication Signup and view all the answers

The cloning vector must have recognition sites for __________ to insert alien DNA efficiently.

restriction enzymes Signup and view all the answers

Match the following recombinant proteins with their therapeutic uses:

Insulin = Diabetes treatment Monoclonal antibodies = Cancer therapy Growth hormone = Growth disorders Erythropoietin = Anemia treatment Signup and view all the answers

Match the following items related to cloning vectors with their descriptions:

Origin of replication (ori) = Controls the replication of linked DNA Selectable marker = Identifies transformants Cloning sites = Recognition for restriction enzymes High copy number = Increases DNA yield per cell Signup and view all the answers

Which of the following is a common selectable marker used in E. coli?

Antibiotic resistance to ampicillin Signup and view all the answers

Single recognition sites for restriction enzymes are preferable in vectors for gene cloning.

True Signup and view all the answers

What is one advantage of cloning DNA with a vector that has a high copy number?

It allows for the recovery of many copies of the target DNA. Signup and view all the answers

What is a method used for directly injecting recombinant DNA into the nucleus of an animal cell?

Micro-injection Signup and view all the answers

What enables bacteria to take up recombinant DNA during transformation?

Heat shock Signup and view all the answers

Agrobacterium tumefaciens is capable of delivering DNA to animal cells.

False Signup and view all the answers

In recombinant DNA technology, the first step is the isolation of DNA.

True Signup and view all the answers

What is the term used to describe bacteria that have been treated to take up DNA?

Competent Signup and view all the answers

Name one enzyme that can be used to break open bacterial cells to release DNA.

Lysozyme Signup and view all the answers

To purify DNA, it needs to be free from other macromolecules such as __________ and proteins.

RNA Signup and view all the answers

The T-DNA delivered by Agrobacterium tumefaciens induces a ______ in the plant cells.

tumor Signup and view all the answers

Match the method of gene transfer with its corresponding descriptor:

Micro-injection = Direct injection of DNA into cell nucleus Biolistics = Bombardment of cells with DNA-coated particles Disarmed pathogen vectors = Using modified viruses to transfer DNA Isolation of DNA = Separation of DNA from other cellular components Signup and view all the answers

Match the following vectors with their primary application:

Ti plasmid = Gene transfer in plants Retrovirus = Gene transfer in animals Plasmid = Cloning in bacteria Bacteriophage = Genetic engineering in bacteria Signup and view all the answers

What method is used to precipitate out purified DNA after its isolation?

Chilled ethanol Signup and view all the answers

Which divalent cation is commonly used to make bacterial cells competent?

Calcium Signup and view all the answers

Recombinant DNA can naturally enter bacterial cells without any treatment.

False Signup and view all the answers

What must occur after bacterial cells are incubated with recombinant DNA on ice and then heat shocked?

Return to ice Signup and view all the answers

Micro-particles used in biolistics must be made of gold or tungsten.

True Signup and view all the answers

Herbert Boyer discovered that restriction enzymes can cut DNA strands in random patterns.

False Signup and view all the answers

What process did Boyer and Cohen enable bacteria to act as in biotechnology?

manufacturing plants for specific proteins Signup and view all the answers

Biotechnology is concerned with using live organisms or _______ from organisms.

enzymes Signup and view all the answers

Which of the following correctly describes plasmids?

Small ringlets of DNA that replicate independently Signup and view all the answers

Making bread is not considered a form of biotechnology.

False Signup and view all the answers

What is the primary focus of biotechnology in its restricted sense today?

Genetically modified organisms Signup and view all the answers

What is the role of the origin of replication in cloning?

It initiates the replication of DNA. Signup and view all the answers

Plasmids are used as vectors to transfer DNA into host organisms.

True Signup and view all the answers

What enzyme is primarily responsible for joining DNA molecules together during the process of forming recombinant DNA?

DNA ligase Signup and view all the answers

The first recombinant DNA was constructed by linking a gene encoding __________ resistance with a plasmid.

antibiotic Signup and view all the answers

Which organism was used to first replicate an antibiotic resistance gene?

Escherichia coli Signup and view all the answers

Cloning only happens when a piece of DNA is isolated from its original source.

False Signup and view all the answers

What is the term used for making multiple identical copies of a template DNA?

cloning Signup and view all the answers

What do restriction endonucleases specifically recognize in DNA?

Palindromic nucleotide sequences Signup and view all the answers

Restriction enzymes always cut DNA at the same position within their recognition sequence.

False Signup and view all the answers

Restriction endonucleases leave __________ stretches at the ends of DNA fragments after cutting.

sticky ends Signup and view all the answers

Match the following sequences with their complementary strands:

5' - GAATTC - 3' = 3' - CTTAAG - 5' 3' - GCTAGC - 5' = 5' - CGATCG - 3' 5' - ATCG - 3' = 3' - TAGC - 5' 3' - AAGCTT - 5' = 5' - TTCGA - 3' Signup and view all the answers

What role does DNA ligase play in recombinant DNA technology?

It facilitates the joining of DNA fragments Signup and view all the answers

To create a recombinant vector molecule, both DNA and vector must be cut with the same restriction enzyme.

True Signup and view all the answers

What is the main advantage of using restriction enzymes in genetic engineering?

Creation of recombinant DNA Signup and view all the answers

Recombinant proteins are produced by expressing foreign genes in their native host organisms.

False Signup and view all the answers

What is a bioreactor used for in biotechnology?

To produce large quantities of desired proteins or products. Signup and view all the answers

The ________ involves optimizing conditions to stimulate the expression of a cloned gene.

expression Signup and view all the answers

Why is it important to produce foreign proteins on a large scale?

To meet industrial and pharmaceutical needs. Signup and view all the answers

Only small-scale cultures can yield appreciable quantities of recombinant proteins.

False Signup and view all the answers

What is the significance of using a continuous culture system?

It maintains cells in their physiologically active log phase for better production. Signup and view all the answers

Which method involves directly injecting recombinant DNA into an animal cell's nucleus?

Micro-injection Signup and view all the answers

Biolistics involves bombarding cells with high-velocity micro-particles coated with DNA.

True Signup and view all the answers

Name one type of enzyme used to isolate DNA from bacterial cells.

lysozyme Signup and view all the answers

The process of introducing recombinant DNA into the host organism is known as __________.

transformation Signup and view all the answers

Match the following components with their respective functions:

Lysozyme = Breaks down bacterial cell walls Ribonuclease = Removes RNA Protease = Removes proteins Ethanol = Precipitates purified DNA Signup and view all the answers

What is the first step in the recombinant DNA technology process?

Isolation of DNA Signup and view all the answers

Chilled ethanol is used to remove lipids from the DNA isolation process.

False Signup and view all the answers

During DNA isolation, proteins are removed by treatment with __________.

protease Signup and view all the answers

What is the primary purpose of using gel electrophoresis in DNA analysis?

To separate DNA fragments based on size Signup and view all the answers

What compound is commonly used to stain DNA fragments for visibility under UV light?

ethidium bromide Signup and view all the answers

DNA fragments are negatively charged and migrate towards the ______ during gel electrophoresis.

anode Signup and view all the answers

Match the components with their functions in gel electrophoresis:

Agarose = Matrix for separation Ethidium bromide = Visualization under UV light Anode = Positive electrode Gel electrophoresis = Technique for separation of DNA Signup and view all the answers

What is the function of elution in the context of gel electrophoresis?

To cut bands from the agarose gel Signup and view all the answers

The smaller the DNA fragment, the farther it moves in gel electrophoresis.

True Signup and view all the answers

What is the significance of the agarose gel in gel electrophoresis?

It acts as a medium for the separation of DNA fragments by size. Signup and view all the answers

Which of the following processes involves the use of DNA ligase?

Ferrying DNA into host organisms Signup and view all the answers

Eukaryotic cells have their own restriction endonucleases.

False Signup and view all the answers

What is the purpose of using a bioreactor in large scale production?

To provide controlled conditions for the growth of microorganisms and production of recombinant proteins. Signup and view all the answers

The __________ enzyme is used to monitor the transformation of host cells by foreign DNA.

reporter Signup and view all the answers

Match the following components of the recombinant DNA process with their functions:

Restriction Endonucleases = Cut DNA at specific sites DNA Ligase = Join DNA fragments together Plasmid Vectors = Transport foreign DNA into host cells Bioreactors = Cultivate cells for protein production Signup and view all the answers

What is the first step in recombinant DNA technology?

Isolation of genetic material Signup and view all the answers

DNA is the only type of nucleic acid that serves as the genetic material in all organisms.

False Signup and view all the answers

What method involves injecting recombinant DNA directly into the nucleus of an animal cell?

Micro-injection Signup and view all the answers

The method known as _______ uses high velocity micro-particles to transfer DNA into plant cells.

biolistics Signup and view all the answers

Match the following enzymes with their function:

Lysozyme = Breaks down bacterial cell walls Cellulase = Digests cellulose in plant cells Ribonuclease = Removes RNA Protease = Degrades proteins Signup and view all the answers

What is the purpose of chilling ethanol during DNA precipitation?

To precipitate pure DNA Signup and view all the answers

The process of culturing host cells occurs after ligation of the DNA fragment into a vector.

True Signup and view all the answers

Which of the following is a core technique of modern biotechnology?

Gene cloning Signup and view all the answers

Sexual reproduction allows for variations in genetic traits compared to asexual reproduction.

True Signup and view all the answers

What is the role of bioprocess engineering in biotechnology?

To maintain a sterile environment for the growth of desired microorganisms. Signup and view all the answers

The definition of biotechnology includes the integration of natural science and organisms, cells, parts thereof, and molecular __________ for products and services.

analogues Signup and view all the answers

Match the following biotechnology terms with their correct descriptions:

Genetic engineering = Altering genetic material to change an organism's phenotype Bioprocess engineering = Maintaining a sterile environment for microbial growth Gene cloning = Isolating and replicating a specific DNA segment Recombinant DNA = Mixing DNA from different sources Signup and view all the answers

Which statement is accurate regarding a piece of DNA transferred into an alien organism?

It is most likely not to multiply itself in the progeny cells. Signup and view all the answers

Traditional hybridisation procedures are more efficient than genetic engineering in isolating desirable genes.

False Signup and view all the answers

What is one of the main advantages of sexual reproduction over asexual reproduction?

It allows for genetic variation. Signup and view all the answers

What characteristic of plasmids affects the number of copies per cell?

Origin of replication Signup and view all the answers

Bacteriophages typically have a low copy number of their genome compared to plasmids within bacterial cells.

False Signup and view all the answers

The cloning vector must ideally have few recognition sites for __________ enzymes to avoid generating multiple fragments.

restriction Signup and view all the answers

Which antibiotic resistance genes are considered useful selectable markers for E. coli?

Both B and C Signup and view all the answers

Multiple recognition sites within a vector are beneficial for gene cloning.

False Signup and view all the answers

Name an example of a selectable marker gene used in E. coli.

Ampicillin resistance gene Signup and view all the answers

The use of bioreactors allows for the large-scale production of desired proteins.

True Signup and view all the answers

What is the term used for a protein that is expressed from a foreign gene in a host organism?

Recombinant protein Signup and view all the answers

Bioreactors can be described as vessels where raw materials are biologically converted into specific ______.

products Signup and view all the answers

Why is it important to produce proteins on a large scale in biotechnology?

To meet industrial demands and research needs Signup and view all the answers

Only small volume cultures can yield appreciable quantities of products.

False Signup and view all the answers

What type of host cells can be used for expressing foreign genes?

Bacterial, plant, or animal cells Signup and view all the answers

What is the primary function of restriction enzymes?

To cut DNA at specific sequences Signup and view all the answers

Exonucleases and endonucleases are the same in function.

False Signup and view all the answers

Name one of the key tools used in recombinant DNA technology.

Restriction enzymes Signup and view all the answers

The first restriction endonuclease identified was _____.

Hind II Signup and view all the answers

Which step comes first in genetically modifying an organism?

Identification of DNA with desirable genes Signup and view all the answers

There are over 900 restriction enzymes that have been characterized so far.

True Signup and view all the answers

What are the two types of nucleases mentioned?

Exonucleases and endonucleases Signup and view all the answers

Study Notes

Herbert Boyer's Life and Work

Born in 1936, raised in western Pennsylvania
Earned graduate degree at University of Pittsburgh (1963)
Completed post-graduate studies at Yale (1963-1966)
Became an assistant professor at University of California, San Francisco in 1966
Conducted research on restriction enzymes in E. coli (1969)
Discovered enzymes' ability to cut DNA at specific sites, creating "sticky ends"
Discovered that these clipped ends made pasting together pieces of DNA a precise exercise.
Collaborated with Stanley Cohen in Hawaii to further research DNA splicing.
Their collaboration established the biotechnology field, creating recombinant DNA technology.
Boyer's graduate studies included the University of Pittsburgh and Yale.
Boyer was a key figure in founding the biotechnology field.

DNA Splicing and Recombinant DNA

Boyer and Cohen combined DNA splicing with plasmid transfer methods, which used plasmids as vectors.
Plasmids are small, circular, self-replicating DNA molecules found in bacteria.
They are used as vectors for DNA transfer, carrying a DNA fragment from one cell to another.
Plasmids can replicate independently and carry foreign DNA.
This allowed for precise insertion of DNA segments into bacterial cells, enabling the manufacture of specific proteins.
This process led to the creation of recombinant DNA molecules, which contain DNA from different sources.
The process involves cutting DNA using restriction enzymes, and then joining using DNA ligase, creating a desired genetic combination.
Plasmids are crucial to replicate in bacteria, as they replicate autonomously, facilitating the transfer of foreign DNA into host cells and subsequent replication.

Genetic Engineering Principles

Genetic engineering involves altering the chemistry of genetic material (DNA and RNA).
Traditional hybridisation methods can lead to undesirable genes being included along with desirable ones; this technique overcomes this issue.
Techniques such as gene cloning, gene transfer, and making recombinant DNA allow for the isolation and introduction of desired genes, without the inclusion of undesirable genes.
This allows for precise manipulation of genes for desired characteristics by isolating specific genes and introducing them.
Genetic engineering can isolate specific genes for insertion, making this technique more controlled than older methods.

DNA Replication in Bacteria

Alien DNA transferred into an organism may not replicate unless integrated into the host DNA.
This depends on integration into host DNA for replication and inheritance in progeny cells.
Plasmids ensure this replication within the bacterial cell's cytoplasm via an 'origin of replication', controlling replication and ensuring copies are made.
Host DNA replicates the new segments, assuring their transmission through generations, replicating the newly added gene.
Plasmids are crucial for replicating in bacteria, due to their autonomous replication.

Restriction Enzymes

Enzymes that cut DNA at specific locations (recognition sequences).
Two types of nucleases (exonucleases and endonucleases).
Exonucleases remove nucleotides from DNA ends.
Endonucleases cut DNA at specific points within the DNA strand.
Recognition sequences are usually palindromes (sequences that read the same backward and forward).
Create "sticky ends" (single-stranded overhangs), facilitating DNA fragment joining (via DNA ligase).
Named after the bacterial strain in which they were discovered (e.g., EcoRI from Escherichia coli).
Used for cutting DNA at specific points, allowing for the insertion of foreign DNA into DNA.
Restriction enzymes recognize specific DNA sequences, cut DNA at precise locations, creating sticky ends crucial for ligation, ensuring correct joining of DNA.
Recognition sequences are specific, ensuring cutting at the correct sites, using precise molecular tools.

Tools of Recombinant DNA Technology

Restriction enzymes, polymerase enzymes (e.g., DNA ligase), vectors (such as plasmids and bacteriophages), and host organisms are critical to recombinant DNA technology.
Gel electrophoresis is used to separate and visualize DNA fragment sizes, isolating the desired fragment.
Vectors (plasmids, bacteriophages) facilitate transfer, replication of foreign DNA within the host organism.
The process provides the essential tools for manipulating DNA, including cutting, separating, joining, and transferring genes to achieve the intended results.
Vectors are essential carriers of DNA fragments during gene transfer.

Polymerase Chain Reaction (PCR)

PCR is a technique to make millions of copies of DNA sequences in vitro using heat-stable DNA polymerase.
Uses heat-stable DNA polymerase (e.g., Taq polymerase) for multiple-cycle amplification of specific DNA sequences.
PCR steps: Denaturation, annealing, and extension.
Key for amplifying specific DNA sequences for further analysis and use, allowing for precise targeting of specific DNA regions.
PCR uses primers to target specific DNA regions for replication, using a thermostable polymerase like Taq.

Obtaining Foreign Gene Product

Recombinant DNA is inserted into a host cell (often bacteria).
Host cells can multiply and produce many copies of the desired protein through the use of vectors.
Purification and processing are essential to isolate the desired product, often requiring purification techniques like chromatography for separation.
Recombinant DNA technology enables production of large quantities of specific proteins, beneficial for humans.
Production demands may require large-scale procedures.

Bioreactors and Downstream Processing

Bioreactors are large vessels culturing cells or microorganisms to produce desired proteins/compounds on a large scale; they provide optimal conditions.
Downstream processing involves separating and purifying the desired product from the bioreactor's culture using methods like filtration, precipitation, or chromatography, ensuring large scale purification.
Quality control is critical for the final formulated product, ensuring safety and efficacy; quality control methods are used to ensure that the product meets prescribed standards.
Large-scale production methods are critical for obtaining proteins, like antibiotics, or hormones, which have huge demand.
Bioreactors maintain optimal conditions for large-scale production; downstream processing ensures a pure product for applications.

Large Scale Production

Biotechnology produces drugs, enzymes, and other proteins.
Bioreactors are used for large-scale production of biomolecules.
This involves industrial-scale procedures allowing large volumes of desired product, increasing yield needed for commercial products.
Large-scale production facilitates commercial product development, improving availability and accessibility.

Recombinant DNA formation

The same restriction enzyme must be used for both the vector and source DNA.
Plasmid DNA acts as vectors, carrying the foreign DNA into the host organism and enabling replication; the origin of replication is critical for this process.
Recombinant DNA is formed by linking foreign DNA to a vector using DNA ligase, creating a specific desired gene combination.

Techniques of DNA introduction into host cell

Methods exist for introducing recombinant DNA into host cells: direct injection (microinjection), bombardment (biolistics, gene gun), and transfer using vectors (plasmids or bacteriophages).
Heat shocking, for instance, makes cells competent for DNA uptake.
Various methods for introducing DNA into cells; each method has specific purposes.
Competent cells are prepared for DNA uptake using methods such as calcium treatment or electroporation.

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Description

Test your knowledge on the significant discoveries and principles of biotechnology, particularly those contributed by Herbert Boyer. This quiz covers concepts such as restriction enzymes, genetic engineering advantages, and bioprocess engineering techniques. Challenge yourself with matching terms and answering key questions in the field of biotechnology.

NCERT - BIOTECHNOLOGY - PRINCIPLES & PROCESSES - 24-25

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Questions and Answers

What significant property do restriction enzymes discovered by Boyer possess?

Herbert Boyer completed his undergraduate studies at Yale.

Who collaborated with Herbert Boyer in the early development of biotechnology?

Biotechnology uses live organisms or __________ from organisms to produce useful products.

Match the following terms with their correct descriptions:

What process did Boyer and Cohen enable bacteria to perform?

Making bread could be considered a form of biotechnology.

In what year did Herbert Boyer become an assistant professor at the University of California at San Francisco?

Which of the following techniques is primarily associated with altering the chemistry of genetic material?

Asexual reproduction provides more genetic variation than sexual reproduction.

What is the main advantage of using genetic engineering over traditional hybridization methods?

Bioprocess engineering maintains a sterile ambiance to enable growth of only the desired ______ in large quantities.

Match the following terms with their corresponding descriptions:

Which organization provided a definition for biotechnology?

Incorporating a piece of DNA into an alien organism guarantees its multiplication in progeny cells.

What is the significant impact of sexual reproduction compared to asexual reproduction on genetic traits?

Which of the following is the first step in genetically modifying an organism?

Restriction enzymes only make cuts at the ends of DNA molecules.

What is the significance of the recognition sequence in restriction enzymes?

The first restriction endonuclease, Hind II, recognizes a specific sequence of _____ base pairs.

Match the following restriction enzymes with their origin:

Which of the following enzymes is responsible for cutting DNA?

Exonucleases can only remove nucleotides from the ends of the DNA.

A major tool in recombinant DNA technology that introduces DNA into the host is called a _____ .

What do restriction endonucleases cut in the DNA?

Restriction enzymes can create blunt ends when they cut DNA.

What are the single-stranded portions at the ends of a cut DNA called?

Restriction endonucleases recognize specific sequences known as __________.

Match the following terms with their definitions:

Which of the following statements about palindromic DNA sequences is true?

Different restriction enzymes can create compatible sticky ends.

When two DNA fragments are cut using the same restriction enzyme, they can be joined together by __________.

What is the primary component used in gel electrophoresis as the matrix?

DNA fragments can be visualized with the naked eye without staining.

What color do stained DNA bands appear after exposure to UV light?

In gel electrophoresis, DNA fragments migrate towards the ______ because they are negatively charged.

Which of the following correctly describes the movement of DNA fragments during gel electrophoresis?

Elution involves cutting the DNA bands from the agarose gel.

What is the purpose of using ethidium bromide in gel electrophoresis?

What is a primary function of the origin of replication (ori) in a cloning vector?

Bacteriophages have a lower copy number of their genome compared to plasmids within bacterial cells.

What is the significance of having a selectable marker in a cloning vector?

The genes encoding resistance to antibiotics such as ampicillin and __________ are commonly used as selectable markers in E. coli.

Which of the following features is NOT required to facilitate cloning into a vector?

Transformation is the process of introducing foreign DNA into a host bacterium.

How does increasing the copy number of a vector benefit cloning procedures?

Match the following cloning vector components with their functions:

What innovation did Herbert Boyer discover regarding DNA strands?

Biotechnology is only applicable to modern genetically modified organisms.

What was the primary breakthrough that Boyer and Cohen achieved together?

Boyer was born in _____ Pennsylvania.

Match the following scientists with their contributions to biotechnology:

What is one of the main applications of biotechnology as defined in the content?

Restriction enzymes are capable of cutting DNA at any random location.

In what year did Herbert Boyer complete his graduate studies?

What do restriction endonucleases primarily cut?

Sticky ends are produced when restriction enzymes cut DNA at the center of the palindrome sites.

Define a palindromic sequence in DNA.

Restriction endonucleases form __________ molecules of DNA composed of DNA from different sources.

Match the restriction enzyme with its recognition sequence:

What is the significance of sticky ends formed after a restriction enzyme cuts DNA?

Restriction enzymes can cut DNA at multiple different sites on the same molecule.

What enzyme is used to join DNA fragments together after they have been cut with restriction enzymes?

What enzyme is responsible for joining the ends of cut DNA molecules in recombinant DNA technology?

An alien piece of DNA can replicate without being part of a chromosome.

What is the term for making multiple identical copies of a DNA template?

The first recombinant DNA was constructed by linking a gene encoding antibiotic resistance with a native __________ of Salmonella typhimurium.

Match the following components with their functions in recombinant DNA technology:

What is referred to as the origin of replication in a chromosome?

Cloning of the antibiotic resistance gene in E. coli was achieved without the need for a vector.

Who were the two scientists that accomplished the construction of the first recombinant DNA?

Which of the following is the first basic step in genetically modifying an organism?

Restriction enzymes only function at specific sites within the DNA.

What is the function of ligases in recombinant DNA technology?

The enzyme EcoRI is derived from the bacteria __________.

Match the following restriction enzymes with their recognition sequences: