DNA Ligase and Cloning Basics
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Questions and Answers

What is the primary function of DNA ligase in the cloning process?

  • It helps to unwind the DNA strands
  • It facilitates the replication of DNA molecules
  • It separates the DNA into smaller fragments
  • It covalently joins DNA fragments together (correct)

    • What does the term 'cloning' refer to in the context of DNA cloning?

  • Introducing a foreign gene into a host organism
  • Creating a genetically modified organism
  • Making several copies of a DNA molecule (correct)
  • Replicating a DNA molecule in a test tube

    • What is a recombinant DNA molecule?

  • A DNA molecule that has been synthesized in a laboratory
  • A DNA molecule that has been cloned from a living organism
  • A DNA molecule that contains genetic material from two different sources (correct)
  • A DNA molecule that has been modified to express a specific protein

    • What is the primary advantage of using plasmids as cloning vectors?

    <p>They can replicate independently of the host cell's genome</p> Signup and view all the answers

    Which of the following is a required component of a plasmid cloning vector?

    <p>An origin of replication (OriC)</p> Signup and view all the answers

    What is the purpose of including a selectable marker gene, such as an antibiotic resistance gene, in a plasmid cloning vector?

    <p>To facilitate the identification and selection of cells that have taken up the vector</p> Signup and view all the answers

    What technique allows the separation of proteins based on their size?

    <p>Gel electrophoresis (SDS-PAGE)</p> Signup and view all the answers

    Which of the following protein tags is a fluorescent protein that can be detected by its fluorescence?

    <p>Green fluorescent protein (GFP)</p> Signup and view all the answers

    What technique involves the use of antibodies to purify a specific protein from a mixture?

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

    Which of the following protein tags allows for purification using beads with bound nickel ions?

    <p>Poly-His</p> Signup and view all the answers

    What technique involves the transfer of separated proteins from a gel to a membrane for detection with antibodies?

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

    Which of the following protein tags can be purified using a peptide corresponding to its amino acid sequence?

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

    What is a protein domain?

    <p>A compact region of the protein's polypeptide chain with defined structure that can fold independently</p> Signup and view all the answers

    What is the purpose of Green Fluorescent Protein (GFP) in proteins?

    <p>To detect the protein</p> Signup and view all the answers

    What are the characteristics of a protein domain?

    <p>It is self-stabilizing, has defined structure, and can fold independently</p> Signup and view all the answers

    What does GST stand for in the context of proteins?

    <p>Glutathione S transferase</p> Signup and view all the answers

    Why are recombinant proteins produced?

    <p>To generate large quantities of specific proteins for research or commercial use</p> Signup and view all the answers

    What is the purpose of a restriction site in the creation of recombinant DNA?

    <p>To cut the DNA fragments and allow for insertion into the plasmid</p> Signup and view all the answers

    What is the purpose of a DNA ligase in the creation of recombinant DNA?

    <p>To close the plasmid and seal the inserted DNA fragment</p> Signup and view all the answers

    What is the purpose of a promoter sequence in an expression vector?

    <p>To drive the expression of the inserted gene</p> Signup and view all the answers

    What is the purpose of a poly-T primer in the reverse transcription of mRNA?

    <p>To hybridize to the poly-A tail of the mRNA and initiate reverse transcription</p> Signup and view all the answers

    Why is it challenging to express human proteins in bacteria?

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

    What is the purpose of protein tagging in recombinant protein expression?

    <p>To add a sequence of amino acids to the protein for easier purification</p> Signup and view all the answers

    What is the primary purpose of DNA cloning?

    <p>Both a and c</p> Signup and view all the answers

    What are restriction endonucleases?

    <p>Enzymes that recognize and cut DNA at specific sequences</p> Signup and view all the answers

    What type of sequences are recognized by restriction endonucleases?

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

    How do restriction endonucleases cut DNA?

    <p>They can cut DNA in both blunt and staggered (off-center) manners</p> Signup and view all the answers

    What is a vector in DNA cloning?

    <p>A carrier of the gene or DNA segment of interest</p> Signup and view all the answers

    What are the products of a staggered (off-center) cut by a restriction endonuclease?

    <p>Sticky or cohesive ends</p> Signup and view all the answers

    Which of the following protein tags allows for purification using beads with bound nickel ions?

    <p>Poly-His</p> Signup and view all the answers

    What technique involves the transfer of separated proteins from a gel to a membrane for detection with antibodies?

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

    Which of the following protein tags can be purified using a peptide corresponding to its amino acid sequence?

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

    What is the purpose of the FLAG tag in protein purification?

    <p>To allow detection using an antibody against the FLAG peptide</p> Signup and view all the answers

    Which of the following protein tags is a fluorescent protein that can be detected by its fluorescence?

    <p>Green fluorescent proteins (GFP)</p> Signup and view all the answers

    What technique allows the separation of proteins based on their size?

    <p>Gel electrophoresis (SDS-PAGE)</p> Signup and view all the answers

    What technique involves the use of antibodies to purify a specific protein from a mixture?

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

    What is the main purpose of DNA cloning?

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

    What is the key function of restriction endonucleases in DNA cloning?

    <p>To recognize and cut the phosphodiester bond at specific DNA sequences</p> Signup and view all the answers

    What type of DNA ends are generated by a staggered (off-center) cut by a restriction endonuclease?

    <p>Sticky/cohesive ends</p> Signup and view all the answers

    What is the defining characteristic of the DNA sequences recognized by restriction endonucleases?

    <p>They are palindromic sequences</p> Signup and view all the answers

    What type of DNA ends are generated by a blunt cut by a restriction endonuclease?

    <p>Blunt ends</p> Signup and view all the answers

    What is the primary advantage of using plasmids as cloning vectors in DNA cloning?

    <p>Plasmids can replicate independently of the host cell's chromosome</p> Signup and view all the answers

    What is the main purpose of including a selectable marker gene, such as an antibiotic resistance gene, in a plasmid cloning vector?

    <p>To allow for the selection of cells that have successfully incorporated the recombinant plasmid</p> Signup and view all the answers

    What is the role of DNA ligase in cloning?

    <p>Catalyzing the formation of phosphodiester bonds between DNA fragments</p> Signup and view all the answers

    Why are plasmids considered good vectors for cloning?

    <p>They can replicate independently of the bacterial genome</p> Signup and view all the answers

    What is the key feature that plasmid cloning vectors must possess?

    <p>Origin of replication allowing independent replication</p> Signup and view all the answers

    What is the purpose of a selectable gene in a plasmid cloning vector?

    <p>To allow for selection of cells containing the vector</p> Signup and view all the answers

    What is recombinant DNA technology part of?

    <p>Genetic engineering</p> Signup and view all the answers

    Which statement best describes a clone?

    <p>Genetically identical individuals derived from a single source</p> Signup and view all the answers

    What is the key step in creating recombinant DNA using vectors?

    <p>Joining a DNA fragment of interest into the vector</p> Signup and view all the answers

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

    <p>Closing the plasmid</p> Signup and view all the answers

    What is the primary function of a poly-T primer in reverse transcription?

    <p>Selecting human mRNA</p> Signup and view all the answers

    Why are eukaryotic systems like yeast preferred for protein expression over bacteria?

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

    What is the main advantage of using protein tagging in recombinant protein expression?

    <p>Facilitating protein purification</p> Signup and view all the answers

    In the context of gene expression vectors, what do ShineDalgarno (SD) sequences function as?

    <p>Ribosomal binding sequences</p> Signup and view all the answers

    What is the primary purpose of utilizing the same restriction endonuclease to cut both DNA fragments during cloning?

    <p>To create fragments with compatible sticky-ends</p> Signup and view all the answers

    Why is plasmid used as a vector in DNA cloning?

    <p>To carry the DNA segment of interest</p> Signup and view all the answers

    What is the primary function of a protein domain?

    <p>To act as a self-contained functional unit within the protein</p> Signup and view all the answers

    What is the main purpose of using a GST-tag in recombinant protein production?

    <p>To purify the recombinant protein</p> Signup and view all the answers

    What is the primary advantage of using a GFP-tag compared to a GST-tag in recombinant protein production?

    <p>GFP-tagged proteins can be detected without purification</p> Signup and view all the answers

    Which of the following is a key characteristic of a protein domain?

    <p>It can fold independently from the rest of the protein</p> Signup and view all the answers

    What is the primary purpose of producing recombinant proteins?

    <p>To generate large quantities of a desired protein</p> Signup and view all the answers

    What is the main advantage of using plasmids as cloning vectors?

    <p>Plasmids can replicate independently within the host cell</p> Signup and view all the answers

    What is the purpose of a promoter sequence in an expression vector?

    <p>To drive the expression of the recombinant gene</p> Signup and view all the answers

    Which of the following is a required component of a plasmid cloning vector?

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

    What is the primary function of DNA ligase in the cloning process?

    <p>To join DNA fragments together</p> Signup and view all the answers

    What is the purpose of including a selectable marker gene, such as an antibiotic resistance gene, in a plasmid cloning vector?

    <p>To enable the selection of host cells that have successfully taken up the recombinant plasmid</p> Signup and view all the answers

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