Recombinant DNA Technology
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What is the main purpose of Recombinant DNA technology?

  • To create genetically modified organisms using natural processes
  • To study the structure of DNA in different organisms
  • To extract DNA from organisms for research purposes
  • To produce DNA via artificial means using protocols like gene cloning and genetic engineering (correct)

    • Which enzymes are essential for constructing Recombinant DNA?

  • RNA Polymerase and Helicase
  • DNA Polymerase and Primase
  • Ligase and Helicase
  • Restriction Enzymes and DNA Ligase (correct)

    • What is the role of a vector in Recombinant DNA technology?

  • To transfer foreign genetic material into another cell (correct)
  • To amplify DNA fragments
  • To cleave DNA at specific base pair sequences
  • To synthesize new DNA strands

    • Why are Type II restriction endonucleases important in Recombinant DNA technology?

    <p>They cut DNA molecules internally at specific base pair sequences</p> Signup and view all the answers

    What is the process that involves the introduction of rDNA into eukaryotic cells?

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

    How are host cells made more permeable to take up exogenous DNA in the process of cell competency?

    <p>Treatment with calcium chloride</p> Signup and view all the answers

    What technology has allowed for the production of human insulin using bacteria as host cells?

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

    What was the method used for cloning and expression of hGH in Chinese hamster ovary (CHO) cells?

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

    What did the use of mammalian cells ensure in the production of recombinant human growth hormone (hGH)?

    <p>Proper post-translational modifications</p> Signup and view all the answers

    What is the process that can occur naturally in some bacteria or be induced in the laboratory, involving the uptake and incorporation of rDNA into the bacterial cell?

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

    What is the source of the first identified restriction endonuclease, EcoRI?

    <p>Escherichia coli</p> Signup and view all the answers

    How do restriction enzymes derive their names?

    <p>From the bacterial species they come from</p> Signup and view all the answers

    What is the function of sticky ends produced by some restriction enzymes?

    <p>Allow fragments to join through complementary base pairing</p> Signup and view all the answers

    How do blunt ends differ from sticky ends in DNA fragments?

    <p>Blunt ends have no overhangs and make ligation less efficient</p> Signup and view all the answers

    How do host cells protect their own DNA from restriction endonucleases?

    <p>Through methylation</p> Signup and view all the answers

    What are vectors used for in genetic engineering?

    <p>To carry and propagate foreign DNA in a host cell</p> Signup and view all the answers

    What type of DNA molecules are plasmids?

    <p>Circular extrachromosomal DNA molecules used as vectors</p> Signup and view all the answers

    Why are bacteriophages used as vectors?

    <p>For their large insert size and ease of screening through plaque formation</p> Signup and view all the answers

    What distinguishes cosmids from other vectors?

    <ul> <li>Cosmids are hybrid vectors that combine plasmid and bacteriophage features, allowing for the cloning of larger DNA fragments.</li> </ul> Signup and view all the answers

    What is the process that involves the introduction of rDNA into eukaryotic cells?

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

    How are host cells made more permeable to take up exogenous DNA in the process of cell competency?

    <p>Treatment with calcium chloride</p> Signup and view all the answers

    What was the method used for cloning and expression of hGH in Chinese hamster ovary (CHO) cells?

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

    What is the role of recombinant DNA technology in insulin production?

    <p>Allows production of human insulin using bacteria as host cells</p> Signup and view all the answers

    What is the source of recombinant human growth hormone (hGH) prior to the development of recombinant DNA technology?

    <p>Extracted from cadaveric pituitary glands</p> Signup and view all the answers

    What is the process that can occur naturally in some bacteria or be induced in the laboratory, involving the uptake and incorporation of rDNA into the bacterial cell?

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

    What technology has allowed for proper post-translational modifications, such as glycosylation, in the production of recombinant human growth hormone (hGH)?

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

    What is the method for making cells competent by creating temporary pores on the cell membrane?

    <p>Heat shock</p> Signup and view all the answers

    What is transferred from one bacterium to another through a viral vector?

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

    What process involves introducing foreign genetic material into cells?

    <p>Insertion into Host</p> Signup and view all the answers

    What is the main purpose of Recombinant DNA technology?

    <p>To transfer foreign genetic material into another cell using vectors</p> Signup and view all the answers

    What are Type II restriction endonucleases important for in Recombinant DNA technology?

    <p>Cutting DNA molecules internally at specific base pair sequences</p> Signup and view all the answers

    What is the function of a vector in Recombinant DNA technology?

    <p>To transfer foreign genetic material into another cell</p> Signup and view all the answers

    What is the role of a host cell in Recombinant DNA technology?

    <p>To carry out the genetic modification</p> Signup and view all the answers

    What is the purpose of screening/identifying host cell colonies in Recombinant DNA technology?

    <p>To identify the specific colony containing the target gene</p> Signup and view all the answers

    What is a vector in the context of Recombinant DNA technology?

    <p>A vehicle to transfer foreign genetic material into another cell</p> Signup and view all the answers

    What is the function of restriction enzymes in Recombinant DNA technology?

    <p>To cut DNA molecules internally at specific base pair sequences</p> Signup and view all the answers

    What is the basic principle of constructing recombinant DNA?

    <p>Joining insert DNA to a cloning vector and transferring it into a host cell</p> Signup and view all the answers

    Why must both source DNA and cloning vector be consistently cut into discrete and reproducible fragments in Recombinant DNA technology?

    <p>To prevent random integration of foreign genes into host cells' genomes</p> Signup and view all the answers

    What is the purpose of transferring each DNA construct into a host cell in Recombinant DNA technology?

    <p>To maintain it and allow for expression of target genes</p> Signup and view all the answers

    What is the maximum insert size of Bacterial Artificial Chromosomes (BACs)?

    <p>300 kb</p> Signup and view all the answers

    What type of DNA molecules are plasmids?

    <p>Circular extrachromosomal DNA molecules</p> Signup and view all the answers

    Why are bacteriophages used as vectors?

    <p>Due to their large insert size and ease of screening through plaque formation</p> Signup and view all the answers

    How do restriction enzymes derive their names?

    <p>From the bacterial species they come from</p> Signup and view all the answers

    What is the function of sticky ends produced by some restriction enzymes?

    <p>Help in DNA cloning by allowing fragments to join through complementary base pairing</p> Signup and view all the answers

    What distinguishes cosmids from other vectors?

    <p>They combine plasmid and bacteriophage features, allowing for the cloning of larger DNA fragments</p> Signup and view all the answers

    What is the source of the first identified restriction endonuclease, EcoRI?

    <p>E. coli</p> Signup and view all the answers

    Study Notes

    • Escherichia coli (E.coli) is the source of the first identified restriction endonuclease, named EcoRI.

    • Restriction enzymes derive their names from the bacterial species they come from.

    • Restriction enzymes work by recognizing and binding to specific palindromic sequences in DNA, cutting between G and A residues.

    • Sticky ends are produced by some restriction enzymes, which leave single-stranded overhangs and help in DNA cloning by allowing fragments to join through complementary base pairing.

    • Blunt ends result when a restriction enzyme cuts both strands at the same position, leaving no overhangs and making ligation less efficient and more complex.

    • Host cells protect their own DNA from restriction endonucleases through methylation.

    • Vectors are DNA molecules used to carry and propagate foreign DNA in a host cell, and have specific sites for insertion of target DNA, origins of replication, selectable markers, and multiple cloning sites.

    • Plasmids are circular extrachromosomal DNA molecules used as vectors, which can be found in bacteria, eukaryotes, and archaea, and are small, easy to detect due to antibiotic resistance genes, and have a limited insert size.

    • Bacteriophages are used as vectors due to their large insert size and ease of screening through plaque formation.

    • Cosmids are hybrid vectors that combine plasmid and bacteriophage features, allowing for the cloning of larger DNA fragments.

    • Bacterial Artificial Chromosomes (BACs) and Yeast Artificial Chromosomes (YACs) are large DNA constructs used for cloning, with BACs having a bacterial origin and a maximum insert size of 300 kb, and YACs having a yeast origin and a maximum insert size of 2000 kb.

    • Recombinant DNA is introduced into a host cell for replication and expression, with bacteria, yeast, and mammalian cells being used depending on the desired product.

    • Escherichia coli (E.coli) is the source of the first identified restriction endonuclease, named EcoRI.

    • Restriction enzymes derive their names from the bacterial species they come from.

    • Restriction enzymes work by recognizing and binding to specific palindromic sequences in DNA, cutting between G and A residues.

    • Sticky ends are produced by some restriction enzymes, which leave single-stranded overhangs and help in DNA cloning by allowing fragments to join through complementary base pairing.

    • Blunt ends result when a restriction enzyme cuts both strands at the same position, leaving no overhangs and making ligation less efficient and more complex.

    • Host cells protect their own DNA from restriction endonucleases through methylation.

    • Vectors are DNA molecules used to carry and propagate foreign DNA in a host cell, and have specific sites for insertion of target DNA, origins of replication, selectable markers, and multiple cloning sites.

    • Plasmids are circular extrachromosomal DNA molecules used as vectors, which can be found in bacteria, eukaryotes, and archaea, and are small, easy to detect due to antibiotic resistance genes, and have a limited insert size.

    • Bacteriophages are used as vectors due to their large insert size and ease of screening through plaque formation.

    • Cosmids are hybrid vectors that combine plasmid and bacteriophage features, allowing for the cloning of larger DNA fragments.

    • Bacterial Artificial Chromosomes (BACs) and Yeast Artificial Chromosomes (YACs) are large DNA constructs used for cloning, with BACs having a bacterial origin and a maximum insert size of 300 kb, and YACs having a yeast origin and a maximum insert size of 2000 kb.

    • Recombinant DNA is introduced into a host cell for replication and expression, with bacteria, yeast, and mammalian cells being used depending on the desired product.

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    Related Documents

    Recombinant DNA Technology Lecture PDF

    Description

    Learn about the fundamentals of recombinant DNA technology, including gene cloning, genetic engineering, and molecular cloning. Understand how DNA can be produced through artificial means and how recombinant DNA is constructed from different sources using restriction enzymes, DNA ligase, and vectors.

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