Genetic Engineering Overview
44 Questions
8 Views

Choose a study mode

Play Quiz
Study Flashcards
Spaced Repetition
Chat to Lesson

Podcast

Play an AI-generated podcast conversation about this lesson

Questions and Answers

What is genetic engineering defined as?

The direct manipulation of an organism's genes including heritable and non-heritable recombinant DNA constructs.

What do genes represent?

Segments of DNA that determine physical characteristics and provide instructions for cellular function.

DNA is a nucleic acid that plays a vital role in building and maintaining the human structure.

True (A)

What is the definition of a tool or technique?

<p>A device or instrument used to carry out a specific function.</p> Signup and view all the answers

Which of these is a product of genetic engineering?

<p>Genetically Modified Organisms (GMO) Crops (B)</p> Signup and view all the answers

Which of these is a basic technique used in genetic engineering?

<p>All of the above (D)</p> Signup and view all the answers

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

<p>They recognize specific DNA sequences and cut the DNA at those locations.</p> Signup and view all the answers

Restriction enzymes are more commonly found in eukaryotes than in prokaryotes.

<p>False (B)</p> Signup and view all the answers

What do restriction enzymes recognize?

<p>Palindromic sequences (D)</p> Signup and view all the answers

The process of gel electrophoresis separates DNA molecules based on their charge.

<p>True (A)</p> Signup and view all the answers

What is a restriction map?

<p>A map that depicts the locations where restriction enzymes cut a particular segment of DNA.</p> Signup and view all the answers

What is nucleic acid hybridization?

<p>The base pairing of single strands of DNA or RNA from two different sources to form a hybrid double helix.</p> Signup and view all the answers

What is a nucleic acid probe?

<p>A segment of single-stranded DNA used in hybridization that has a predetermined identity.</p> Signup and view all the answers

What is the difference between a Southern blot and a Northern blot?

<p>A Southern blot analyzes DNA in the gel, while a Northern blot analyzes RNA.</p> Signup and view all the answers

What is molecular cloning?

<p>The isolation and incorporation of a piece of DNA into a vector for replication and manipulation.</p> Signup and view all the answers

What are the three main steps of gene cloning?

<p>All of the above (D)</p> Signup and view all the answers

DNA ligase is an enzyme that joins two DNA molecules.

<p>True (A)</p> Signup and view all the answers

What is the purpose of a gene library?

<p>A mixture of cells that contain a variety of genes.</p> Signup and view all the answers

Transformation is commonly used to introduce recombinant DNA into a host organism.

<p>True (A)</p> Signup and view all the answers

How do scientists detect the correct clone?

<p>Initial screening methods like antibiotic resistance or plaque formation are often used.</p> Signup and view all the answers

What are conventional mutagens?

<p>Agents that produce random mutations in DNA.</p> Signup and view all the answers

What is site-directed mutagenesis?

<p>A technique that introduces mutations at a specific location in DNA.</p> Signup and view all the answers

Cassette mutagenesis involves replacing a DNA fragment with a synthetic DNA fragment.

<p>True (A)</p> Signup and view all the answers

What is gene disruption?

<p>The insertion of a cassette into the middle of a gene.</p> Signup and view all the answers

Gene disruption can cause knockout mutations.

<p>True (A)</p> Signup and view all the answers

What are plasmids?

<p>Small, circular DNA molecules found in bacteria (C)</p> Signup and view all the answers

Plasmids can be used as cloning vectors due to their small size, which makes isolating DNA easier.

<p>True (A)</p> Signup and view all the answers

Plasmids have a single origin of replication, limiting the number of copies produced.

<p>False (B)</p> Signup and view all the answers

What is the purpose of selectable markers in plasmids?

<p>They allow scientists to select for bacterial cells containing the plasmid with the cloned gene.</p> Signup and view all the answers

What are the two main methods for transferring vectors into bacterial cells?

<p>A &amp; B (D)</p> Signup and view all the answers

In blue/white screening, what do blue colonies represent?

<p>Bacterial colonies that do not contain the vector with foreign DNA inserted into them.</p> Signup and view all the answers

The lacZ gene is inactivated when foreign DNA is inserted into it, leading to insertional inactivation.

<p>True (A)</p> Signup and view all the answers

Why does the blue color not develop in colonies where the lacZ gene has been inactivated due to foreign DNA insertion?

<p>Because the inactivated lacZ gene cannot process the X-gal substrate, which is necessary for producing the blue color.</p> Signup and view all the answers

What are some characteristics of ideal hosts for cloning vectors?

<p>All of the above (E)</p> Signup and view all the answers

Which of the following organisms are commonly employed as hosts for cloning vectors?

<p>All of the above (D)</p> Signup and view all the answers

Bacillus subtilis is known for its ability to naturally secrete proteins.

<p>True (A)</p> Signup and view all the answers

What are reporter genes?

<p>Genes that encode proteins that are easy to detect and assay.</p> Signup and view all the answers

Which of these are examples of reporter genes?

<p>All of the above (D)</p> Signup and view all the answers

What is the purpose of gene fusions in genetic engineering?

<p>To combine promoters or coding sequences of genes of interest with those of reporter genes, allowing scientists to understand how gene regulation works under different conditions.</p> Signup and view all the answers

MRNA produced from cloned genes must be efficiently translated to produce functional proteins.

<p>True (A)</p> Signup and view all the answers

What are some challenges associated with efficient translation of mRNA?

<p>All of the above (D)</p> Signup and view all the answers

What is the purpose of expression vectors in genetic engineering?

<p>To allow researchers to control the expression of cloned genes, ensuring high levels of protein production through strong promoters and effective termination signals.</p> Signup and view all the answers

Expression vectors are essential for efficient gene expression because they contain strong promoters that enhance the transcription of the cloned gene.

<p>True (A)</p> Signup and view all the answers

Effective transcription terminators prevent the expression of other genes on the plasmid, ensuring controlled gene expression.

<p>True (A)</p> Signup and view all the answers

Flashcards

Genetic Engineering

Direct manipulation of an organism's genes, including heritable and non-heritable recombinant DNA constructs.

Genes

Segments of DNA that determine physical traits and provide instructions for cell function.

DNA

Nucleic acid that carries instructions for building and maintaining the body.

Restriction Enzyme

Enzyme that cuts DNA at specific sequences.

Signup and view all the flashcards

Gel Electrophoresis

Technique that separates DNA fragments by size.

Signup and view all the flashcards

Nucleic Acid Hybridization

Base pairing of single-stranded DNA or RNA from different sources.

Signup and view all the flashcards

Southern Blot

Hybridization procedure where DNA is on the gel and probe is RNA or DNA.

Signup and view all the flashcards

Molecular Cloning

Copying genes by incorporating them into a vector for replication.

Signup and view all the flashcards

Cloning Vector

Carrier for DNA fragment replication.

Signup and view all the flashcards

DNA Ligase

Enzyme that joins DNA molecules together.

Signup and view all the flashcards

Transformation

Introducing recombinant DNA into a host organism.

Signup and view all the flashcards

Gene Library

Collection of cells containing various genes from a source.

Signup and view all the flashcards

Shotgun Cloning

Randomly inserting genome fragments into vectors.

Signup and view all the flashcards

Site-Directed Mutagenesis

Precisely changing a gene sequence in a DNA segment.

Signup and view all the flashcards

Cassette Mutagenesis

Replacing a DNA fragment with another in a specific location.

Signup and view all the flashcards

Knockout Mutations

Disrupting a gene's function by inserting a sequence.

Signup and view all the flashcards

Plasmid

Small, circular DNA molecule that replicates independently in bacteria.

Signup and view all the flashcards

Blue/White Screening

Method used in cloning to visually identify cells containing recombinant DNA.

Signup and view all the flashcards

Insertional Inactivation

Disrupting a gene's function by inserting foreign DNA.

Signup and view all the flashcards

Expression Vectors

Vectors that control the expression of cloned genes.

Signup and view all the flashcards

GMO (Genetically Modified Organisms)

Organisms with altered genes.

Signup and view all the flashcards

Prokaryotes

Single-celled organisms without a nucleus.

Signup and view all the flashcards

Eukaryotes

Single-celled or multi-celled organisms with a nucleus.

Signup and view all the flashcards

Modification Enzymes

Enzymes that protect a cell's DNA from restriction enzymes.

Signup and view all the flashcards

Study Notes

Genetic Engineering Definition

  • Genetic engineering is the direct manipulation of an organism's genes, including heritable and non-heritable recombinant DNA constructs.

Genes vs. DNA

  • Genes are segments of DNA.
  • DNA gives physical characteristics and carries instructions for cell function and growth.
  • DNA is a nucleic acid responsible for building and maintaining human structure.

Tools/Techniques

  • A tool or technique is a device or instrument used to carry out a specific function.

Products of Genetic Engineering

  • Genetically Modified Organisms (GMO) crops include apples, mangoes, seedless grapes, canola oil, and corn oil.

Restriction and Modification Enzymes

  • Genetic engineering uses in vitro techniques to alter genetic material.
  • Basic techniques include restriction enzymes, gel electrophoresis, nucleic acid hybridization, nucleic acid probes, molecular cloning, and cloning vectors.
  • Restriction enzymes recognize specific DNA sequences and cut DNA at those sites.
  • These enzymes are widespread in prokaryotes, but rare in eukaryotes.
  • Restriction enzymes protect prokaryotes from foreign DNA (like viruses).
  • They are essential for in vitro DNA manipulation.
  • Three classes of restriction enzymes: Type II cleave DNA within their recognition sequences.
    • Restriction enzymes recognize inverted repeat sequences (palindromes).
    • These sequences are typically 4-8 base pairs long; EcoRI recognizes a 6-base pair sequence.
    • Restriction enzymes can create either sticky ends or blunt ends.
  • Restriction enzymes protect cells from foreign DNA by destroying it.
  • They also protect their own DNA from accidental destruction.
  • Modification enzymes protect a cell's DNA for restriction enzymes by chemically modifying nucleotides in the restriction recognition sequence.
  • Modification generally involves DNA methylation.

Gel Electrophoresis

  • Gel electrophoresis separates DNA molecules based on size.
  • Electrophoresis uses an electrical field to separate charged molecules.
  • Gels are typically made of agarose, a polysaccharide.
  • Nucleic acids migrate through the gel towards the positive electrode due to their negatively charged phosphate groups.
  • Gels can be stained with ethidium bromide to visualize DNA under UV light.

Determining Fragment Size

  • The same DNA cut with different restriction enzymes will have different banding patterns on an agarose gel.
  • Fragment size can be determined by comparing to a standard.
  • A restriction map is a map showing the location of restriction enzyme cuts on a DNA segment.

Nucleic Acid Hybridization

  • Nucleic acid hybridization is the base pairing of single DNA or RNA strands from different sources to form a hybrid double helix.
  • A segment of single-stranded DNA used in hybridization is called a nucleic acid probe.
  • Southern blotting is a hybridization procedure where DNA is in the gel and the probe is either RNA or DNA.
  • Northern blotting is a similar procedure where RNA is in the gel.

Molecular Cloning

  • Molecular cloning is the process of isolating and incorporating a piece of DNA into a vector for replication and manipulation.
  • Three main steps include:
    • Isolation and fragmentation of source DNA.
    • Insertion of DNA fragment into a cloning vector.
    • Introduction of cloned DNA into a host organism.

Steps in Cloning

  • Step 1: Source DNA is isolated and fragmented.
    • This DNA can be genomic DNA, RNA, or amplified using PCR.
    • Genomic DNA must first be restriction digested.
  • Step 2: The DNA fragment is inserted into a cloning vector (often a plasmid or a virus).
    • Most cloning vectors come from plasmids or viruses.
    • DNA is usually inserted in vitro.
    • DNA ligase joins the DNA molecules together (using sticky or blunt ends).
  • Step 3: The recombinant vector (containing the cloned DNA) is introduced into a host organism.
    • Transformation is often used to introduce recombinant DNA into the host.
    • Some cells contain the desired cloned gene, while others have other genes.
    • A gene library is a collection of cells with multiple genes.
    • Shotgun cloning uses random genome fragments to create gene libraries.

Cloning Detection

  • A common screening method detects correct clones using antibiotic resistance and plaque formation.
  • These can be sufficient for cloning PCR-generated DNA sequences.
  • A heterogeneous gene library might require more detailed analysis.

Molecular Mutagenesis

  • Conventional mutagens produce random mutations.
  • Site-directed mutagenesis performs mutations in vitro at a precise location.
    • Used to study amino acid activity.
    • Structural biologists greatly benefit from this tool.
  • Cassette mutagenesis replaces DNA fragments with synthetic fragments causing, when inserted into gene sequences, "knockouts"; gene disruptions.

Plasmids as Cloning Vectors

  • Plasmids are natural vectors with useful properties for cloning vectors.

    • Small size for easy isolation.
    • Independent replication origin.
    • Multiple copies per cell.
    • Selectable markers are present.

  • Vector transfer is via chemical transformation or electroporation.

  • Blue/white screening identifies clones with or without foreign DNA inserted (insertional inactivation) on vectors.

    • Blue colonies don't have foreign DNA.
    • White colonies have foreign DNA inserted.

Cloning Hosts

  • Ideal hosts are quick-growing, inexpensive, non-pathogenic, and adept at DNA incorporation and replication.
  • Hosts must also be genetically stable.
  • Common hosts are Escherichia coli, Bacillus subtilis, and Saccharomyces cerevisiae.

Shuttle Vectors

  • Shuttle vectors enable efficient eukaryotic mRNA translation and gene expression.
  • Shuttle vectors solve problems by using promoters, terminators, and codon usage to express cloned genes with high levels of protein in various organisms.

Studying That Suits You

Use AI to generate personalized quizzes and flashcards to suit your learning preferences.

Quiz Team

Related Documents

Biotech Q3 Exam Review PDF

Description

Explore the fundamentals of genetic engineering, including the definition of genes and DNA, key tools and techniques, and products like GMOs. This quiz delves into the methods used in manipulating genetic materials and their implications in modern science.

More Like This

Genetic Engineering Overview
40 questions

Genetic Engineering Overview

HandsDownAnemone avatar
HandsDownAnemone
Genetic Engineering Overview
33 questions

Genetic Engineering Overview

FriendlyTrust avatar
FriendlyTrust
Genetic Engineering and GMOs Quiz
15 questions

Genetic Engineering and GMOs Quiz

IntuitiveClarity2296 avatar
IntuitiveClarity2296
Recombinant DNA Quiz
45 questions

Recombinant DNA Quiz

FerventGamelan avatar
FerventGamelan
Use Quizgecko on...
Browser
Open
Browser
Browser