DNA Ligase and Cloning Basics

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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?

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

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

An origin of replication (OriC) 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?

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

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

Gel electrophoresis (SDS-PAGE) Signup and view all the answers

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

Green fluorescent protein (GFP) Signup and view all the answers

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

Immunoprecipitation Signup and view all the answers

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

Poly-His Signup and view all the answers

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

Immunoblotting Signup and view all the answers

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

All of the above Signup and view all the answers

What is a protein domain?

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

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

To detect the protein Signup and view all the answers

What are the characteristics of a protein domain?

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

What does GST stand for in the context of proteins?

Glutathione S transferase Signup and view all the answers

Why are recombinant proteins produced?

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

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

To cut the DNA fragments and allow for insertion into the plasmid Signup and view all the answers

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

To close the plasmid and seal the inserted DNA fragment Signup and view all the answers

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

To drive the expression of the inserted gene Signup and view all the answers

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

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

Why is it challenging to express human proteins in bacteria?

All of the above Signup and view all the answers

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

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

What is the primary purpose of DNA cloning?

Both a and c Signup and view all the answers

What are restriction endonucleases?

Enzymes that recognize and cut DNA at specific sequences Signup and view all the answers

What type of sequences are recognized by restriction endonucleases?

Palindromic sequences Signup and view all the answers

How do restriction endonucleases cut DNA?

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

What is a vector in DNA cloning?

A carrier of the gene or DNA segment of interest Signup and view all the answers

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

Sticky or cohesive ends Signup and view all the answers

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

Poly-His Signup and view all the answers

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

Immunoblotting Signup and view all the answers

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

HA Signup and view all the answers

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

To allow detection using an antibody against the FLAG peptide Signup and view all the answers

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

Green fluorescent proteins (GFP) Signup and view all the answers

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

Gel electrophoresis (SDS-PAGE) Signup and view all the answers

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

Immunoprecipitation Signup and view all the answers

What is the main purpose of DNA cloning?

All of the above Signup and view all the answers

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

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

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

Sticky/cohesive ends Signup and view all the answers

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

They are palindromic sequences Signup and view all the answers

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

Blunt ends Signup and view all the answers

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

Plasmids can replicate independently of the host cell's chromosome 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?

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

What is the role of DNA ligase in cloning?

Catalyzing the formation of phosphodiester bonds between DNA fragments Signup and view all the answers

Why are plasmids considered good vectors for cloning?

They can replicate independently of the bacterial genome Signup and view all the answers

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

Origin of replication allowing independent replication Signup and view all the answers

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

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

What is recombinant DNA technology part of?

Genetic engineering Signup and view all the answers

Which statement best describes a clone?

Genetically identical individuals derived from a single source Signup and view all the answers

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

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

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

Closing the plasmid Signup and view all the answers

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

Selecting human mRNA Signup and view all the answers

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

To prevent post-translational modifications Signup and view all the answers

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

Facilitating protein purification Signup and view all the answers

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

Ribosomal binding sequences Signup and view all the answers

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

To create fragments with compatible sticky-ends Signup and view all the answers

Why is plasmid used as a vector in DNA cloning?

To carry the DNA segment of interest Signup and view all the answers

What is the primary function of a protein domain?

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

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

To purify the recombinant protein 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?

GFP-tagged proteins can be detected without purification Signup and view all the answers

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

It can fold independently from the rest of the protein Signup and view all the answers

What is the primary purpose of producing recombinant proteins?

To generate large quantities of a desired protein Signup and view all the answers

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

Plasmids can replicate independently within the host cell Signup and view all the answers

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

To drive the expression of the recombinant gene Signup and view all the answers

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

All of the above Signup and view all the answers

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

To join DNA fragments together 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?

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

DNA Ligase and Cloning Basics

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Questions and Answers

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

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

What is a recombinant DNA molecule?

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

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

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

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

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

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

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

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

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

What is a protein domain?

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

What are the characteristics of a protein domain?

What does GST stand for in the context of proteins?

Why are recombinant proteins produced?

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

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

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

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

Why is it challenging to express human proteins in bacteria?

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

What is the primary purpose of DNA cloning?

What are restriction endonucleases?

What type of sequences are recognized by restriction endonucleases?

How do restriction endonucleases cut DNA?

What is a vector in DNA cloning?

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

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

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

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

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

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

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

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

What is the main purpose of DNA cloning?

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

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

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

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

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

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

What is the role of DNA ligase in cloning?

Why are plasmids considered good vectors for cloning?

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

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

What is recombinant DNA technology part of?

Which statement best describes a clone?

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

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

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

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

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

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

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

Why is plasmid used as a vector in DNA cloning?

What is the primary function of a protein domain?

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

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

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

What is the primary purpose of producing recombinant proteins?

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

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

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

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

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

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