Gene Cloning
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Questions and Answers

What is the purpose of the strong promoter in an expression vector?

  • To inhibit transcription of the gene of interest
  • To enhance translation of the gene of interest
  • To drive high-level transcription of the gene of interest (correct)
  • To prevent translation of the gene of interest

Why is a transcription termination sequence necessary in an expression vector?

  • To initiate transcription of the gene of interest
  • To stop transcription at the appropriate point (correct)
  • To stabilize the plasmid vector
  • To enhance transcription of the gene of interest

What is the role of the antibiotic resistance sequences in the expression vector?

  • To enhance the expression of the gene of interest
  • To allow for selection of transformed cells containing the plasmid (correct)
  • To inhibit the growth of transformed cells
  • To facilitate the insertion of the gene of interest

Why is the blue/white screening method used in identifying recombinant plasmids?

<p>To distinguish between plasmids with and without the gene of interest (D)</p> Signup and view all the answers

What is the purpose of the radioactive DNA probe in the identification of recombinant plasmids?

<p>To specifically bind to the gene of interest (A)</p> Signup and view all the answers

Why is a ribosomal binding site and start codon necessary in an expression vector?

<p>To initiate translation of the gene of interest (A)</p> Signup and view all the answers

What is the function of the polylinker in an expression vector?

<p>To facilitate the insertion of the gene of interest (C)</p> Signup and view all the answers

Why are transformed bacterial cells grown on medium containing ampicillin during the identification process?

<p>To select for cells containing the plasmid (D)</p> Signup and view all the answers

What is gene cloning?

<p>Insertion of foreign DNA into a replicating cell to produce many copies of the DNA (B)</p> Signup and view all the answers

What is the purpose of Southern blot technique in gene cloning?

<p>To locate a specific gene of interest (D)</p> Signup and view all the answers

What is the function of an expression vector in gene cloning?

<p>To introduce a specific gene into a target cell for protein expression (B)</p> Signup and view all the answers

What is the purpose of using a plasmid in gene cloning?

<p>To act as an expression vector for protein production (C)</p> Signup and view all the answers

What is the role of regulatory sequences in an expression vector?

<p>To enhance and promote efficient transcription and translation of the gene (A)</p> Signup and view all the answers

What is the first step in the gene cloning process?

<p>Extract and purify DNA from cells (B)</p> Signup and view all the answers

What is the purpose of cutting DNA with restriction enzymes in gene cloning?

<p>To obtain DNA fragments for further manipulation (C)</p> Signup and view all the answers

What is the function of Agarose gel electrophoresis in gene cloning?

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

Flashcards

Strong Promoter

A DNA sequence within an expression vector that drives strong transcription of the gene of interest.

Transcription Termination Sequence

A sequence within an expression vector that signals the end of transcription, ensuring the proper termination of the RNA transcript.

Antibiotic Resistance Sequences

Sequences within an expression vector that confer resistance to a specific antibiotic, allowing for selection of cells that have successfully incorporated the plasmid.

Blue/White Screening

A technique used to identify recombinant plasmids, based on the ability of bacteria containing the plasmid to produce a blue or white colony on a specific culture medium.

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Plasmid

A small, circular piece of DNA that serves as a carrier for foreign DNA in gene cloning.

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Gene Cloning

A technique involving the insertion of foreign DNA into a replicating cell, allowing for the production of many copies of that DNA.

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Expression Vector

A specialized vector designed to express a specific gene within a target cell, leading to the production of the encoded protein.

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Regulatory Sequences

A set of DNA sequences located upstream of the gene of interest within an expression vector, responsible for regulating the efficiency of gene expression.

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Polylinker

A specific DNA sequence within an expression vector where foreign DNA can be inserted.

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Southern Blot

A technique used in gene cloning to identify specific genes of interest within a sample of DNA.

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Ribosomal Binding Site

A sequence within an expression vector that allows the ribosome to bind and initiate translation of the gene of interest.

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Start Codon

A specific DNA sequence within an expression vector that signifies the start of translation.

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Agarose Gel Electrophoresis

A technique used in gene cloning to separate DNA fragments based on their size.

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Radioactive DNA Probe

A radioactive DNA probe used in gene cloning to selectively bind to the gene of interest.

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DNA Extraction

The process of isolating and purifying DNA from cells as a first step in gene cloning.

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Restriction Enzyme Digestion

The process of cutting DNA using restriction enzymes to create specific fragments for further manipulation in gene cloning.

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