Recombinant DNA Technology Overview
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Questions and Answers

Who developed the method for removing plasmids from cells?

  • Boyer
  • In vitro
  • EFB
  • Cohen (correct)

    • What breakthrough enabled the recombination of segments of DNA?

    DNA splicing

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

    European Federation of Biotechnology

    Biotechnology only includes the processes involving genetically modified organisms.

    <p>False</p> Signup and view all the answers

    Match the following biotechnological processes with their descriptions:

    <p>In vitro fertilisation = Creating a 'test-tube' baby Gene synthesis = Creating synthetic genes Biotechnology = Integration of science and organisms DNA vaccine development = Using DNA to induce immunity</p> Signup and view all the answers

    What does biotechnology primarily deal with?

    <p>Using live organisms or enzymes from organisms</p> Signup and view all the answers

    Curd and wine production can be considered forms of biotechnology.

    <p>True</p> Signup and view all the answers

    What is one application of biotechnology mentioned in producing specific proteins?

    <p>Using bacterial cells as manufacturing plants</p> Signup and view all the answers

    What is the first step in genetically modifying an organism?

    <p>Identification of DNA with desirable genes</p> Signup and view all the answers

    Exonucleases make cuts at specific positions within the DNA.

    <p>False</p> Signup and view all the answers

    The first restriction endonuclease discovered was called _____ II.

    <p>Hind</p> Signup and view all the answers

    Which gene is inactivated due to the insertion of alien DNA?

    <p>β-galactosidase gene</p> Signup and view all the answers

    Agrobacterium tumefaciens delivers a piece of DNA known as '____________' to transform plant cells.

    <p>T-DNA</p> Signup and view all the answers

    Which of the following is NOT a tool of recombinant DNA technology?

    <p>Microbial fermentation</p> Signup and view all the answers

    Recombinant colonies can produce blue colored colonies when the plasmid has an insert.

    <p>False</p> Signup and view all the answers

    What type of substrate is used to differentiate recombinants from non-recombinants?

    <p>Chromogenic substrate</p> Signup and view all the answers

    What is the primary component of nucleic acids that serves as genetic material in most organisms?

    <p>Deoxyribonucleic acid (DNA)</p> Signup and view all the answers

    Restriction enzymes are used to cut RNA at specific locations.

    <p>False</p> Signup and view all the answers

    What is the purpose of using alternative selectable markers in genetic engineering?

    <p>To differentiate recombinants and non-recombinants</p> Signup and view all the answers

    In what year were the two enzymes responsible for restricting the growth of bacteriophage in Escherichia coli isolated?

    <p>1963</p> Signup and view all the answers

    The process of inserting recombinant DNA into bacterial cells is straightforward and does not require any special techniques.

    <p>False</p> Signup and view all the answers

    EcoRI is named after Escherichia coli RY 13.

    <p>True</p> Signup and view all the answers

    What are the blue colored colonies indicative of in the context of recombinant DNA?

    <p>Non-recombinant colonies</p> Signup and view all the answers

    How many different restriction enzymes are known today?

    <p>More than 900</p> Signup and view all the answers

    Match the following terms with their definitions:

    <p>Restriction enzymes = Enzymes that cut DNA at specific sequences Endonucleases = Cut DNA within the strand Exonucleases = Remove nucleotides from the DNA ends Vectors = Carry DNA into host cells</p> Signup and view all the answers

    Match the following organisms with their ability to deliver genes:

    <p>Agrobacterium tumefaciens = Transmits T-DNA to plants Retroviruses = Transforms normal cells into cancerous cells Bacteria = Transferring genes to eukaryotic cells Viruses = Forcing host cells to produce required chemicals</p> Signup and view all the answers

    What type of enzyme is used to remove proteins during the DNA isolation process?

    <p>Protease</p> Signup and view all the answers

    To release DNA from bacterial cells, one can use the enzyme _____ to break the cell wall.

    <p>Lysozyme</p> Signup and view all the answers

    Match the following enzymes with their respective function:

    <p>Lysozyme = Breaks down bacterial cell walls Ribonuclease = Removes RNA Protease = Degrades proteins Ligase = Joins DNA fragments</p> Signup and view all the answers

    What is the purpose of agarose gel electrophoresis in DNA manipulation?

    <p>To separate DNA fragments based on size</p> Signup and view all the answers

    Ethanol is added to a DNA solution to increase its solubility.

    <p>False</p> Signup and view all the answers

    What is the end product formed when the 'gene of interest' is joined with the vector DNA?

    <p>Recombinant DNA</p> Signup and view all the answers

    What is the purpose of PCR in biotechnology?

    <p>To amplify specific segments of DNA</p> Signup and view all the answers

    The enzyme DNA polymerase used in PCR is not thermally stable.

    <p>False</p> Signup and view all the answers

    Name a method used to introduce recombinant DNA into host cells.

    <p>Transformation</p> Signup and view all the answers

    The enzyme _____ is used to extend primers in PCR.

    <p>DNA polymerase</p> Signup and view all the answers

    Match each term with its correct description:

    <p>Thermus aquaticus = Source of thermostable DNA polymerase Selectable marker = Gene allowing resistance to antibiotics Agar plates = Medium for selecting transformed cells Recombinant DNA = DNA formed by combining different sources</p> Signup and view all the answers

    During which step of PCR do primers bind to the template DNA?

    <p>Primer annealing</p> Signup and view all the answers

    Inserting an antibiotic resistance gene into E. coli allows for the differentiation of transformed cells.

    <p>True</p> Signup and view all the answers

    What is the main goal of introducing foreign DNA into host cells?

    <p>To produce a desirable protein</p> Signup and view all the answers

    Study Notes

    Recombinant DNA Technology

    • Boyer and Cohen combined DNA splicing with Cohen's method of removing and reinserting plasmids into bacterial cells, resulting in the ability to recombine DNA segments and insert them into bacteria, making them act as protein factories.
    • This technique is the cornerstone of biotechnology, which uses organisms or enzymes to create useful products and processes.
    • The European Federation of Biotechnology defines biotechnology as the integration of natural science and organisms, cells, parts thereof, and molecular analogues for products and services.

    Tools of Recombinant DNA Technology

    • Restriction endonucleases are enzymes that recognize specific DNA sequences and cut the DNA molecule at those sites.
    • Hind II was the first restriction endonuclease discovered, recognizing a specific sequence of 6 base pairs.
    • Over 900 restriction enzymes, recognizing various sequences, have been isolated from over 230 strains of bacteria.
    • Restriction enzymes are a type of nucleases, which can be exonucleases (removing nucleotides from DNA ends) or endonucleases (cutting within DNA molecules at specific points).

    Processes of Recombinant DNA Technology

    • Gene cloning involves identifying a desired gene, introducing it into a host organism, and ensuring its maintenance and transfer to the host’s progeny.

    Isolation of Genetic Material (DNA)

    • To extract DNA, cells are treated with enzymes like lysozyme (bacteria), cellulase (plant cells), and chitinase (fungus) to break open the cell membrane.
    • Proteins are removed using protease, and ribonuclease eliminates RNA.
    • DNA precipitates out after chilled ethanol is added, appearing as fine threads.
    • Agarose gel electrophoresis is used to separate DNA molecules based on size.

    Cutting DNA at Specific Locations

    • Restriction enzyme digestion is performed by incubating purified DNA with the appropriate restriction enzyme under optimal conditions.
    • Vector DNA is also cut with the same restriction enzyme to create complementary ends for the target DNA fragment.

    Joining DNA Fragments

    • The cut target DNA and vector DNA are mixed, and ligase enzyme is added to join the fragments, creating recombinant DNA.

    Amplifying Gene of Interest

    • Polymerase Chain Reaction (PCR) is used to amplify DNA segments in vitro.
    • PCR uses primers (oligonucleotides complementary to the target DNA), DNA polymerase, and nucleotides.
    • The process involves three steps: denaturation, primer annealing, and extension.
    • Thermostable DNA polymerase (from Thermus aquaticus) is used in PCR to withstand the high temperatures needed for denaturation.

    Inserting Recombinant DNA into the Host Cell

    • Transformation introduces recombinant DNA into recipient cells.
    • Competent cells are made capable of taking up DNA from their surroundings.
    • Selectable markers, like antibiotic resistance genes, are used to identify transformed cells.
    • Insertional inactivation uses a chromogenic substrate to differentiate recombinants from non-recombinants based on their ability to produce color.

    Obtaining the Foreign Gene Product

    • When recombinant DNA is introduced into a host cell, the foreign gene is multiplied.
    • The ultimate goal of recombinant technology is usually to produce a specific protein.
    • Expression of the foreign gene under appropriate conditions leads to the production of the desired protein.

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    Description

    Explore the principles and tools of recombinant DNA technology, including its historical context and significance in biotechnology. Learn about key components such as restriction endonucleases and their role in DNA manipulation.

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