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Exam 20: Recombinant DNA Technology

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Question 21

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In recombinant DNA technology,YAC,RFLP,and λ have identical uses.

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Question 22

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Assume that a given plasmid vector to be used in a cloning experiment contains 4000 base pairs of DNA.Assume also that the restriction endonuclease Cuj cuts this plasmid at the following sites (starting from an arbitrary zero point): 1000,1500,and 3000.Given complete digestion of the plasmid with the endonuclease so that only linear fragments are produced,what sizes of DNA are expected?

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500 bp,150...

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Question 23

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Following are four processes common to most cloning experiments: a) transforming bacteria b) plating bacteria on selective medium c) cutting DNA with restriction endonucleases d) ligating DNA fragments Place components of this list in the order in which they would most likely occur during a cloning experiment.

Some restriction endonucleases are capable of producing blunt ends; others can generate "sticky" ends.

In the context of molecular genetics,reverse transcription PCR (RT-PCR) refers to ________.

What might be a reasonable function of restriction endonucleases in a bacterium,distinct from their use by molecular biologists?

To isolate a bacterium with a plasmid that carries a desired DNA fragment cloned within the ampicillin resistance gene,we should grow bacteria in a medium that contains ampicillin.

When propagating a clone in the lambda phage,would you have more immediate success if the phage entered the lysogenic or the lytic cycle?

Nucleic acid blotting is commonly used in molecular biology.Two types,Southern blots and northern blots,involve gel electrophoresis and a filter,which holds the nucleic acid.Briefly describe the procedure of "blotting" in this context and differentiate between Southern and northern blots.

A knockout animal,in the context of recombinant DNA technology,is one that has been completely anesthetized.

Nucleic acid blotting is widely used in recombinant DNA technology.In a Southern blot,one generally ________.

In recombinant DNA technology,a YAC is an enzyme isolated from a large South American,four-legged mammal.

Restriction endonucleases are especially useful if they generate "sticky" ends.What makes an end sticky?

E.coli is a common YAC.

If one wishes to clone a gene using typical restriction endonucleases,how does the restriction endonuclease recognize genes in the genome?

A restriction map provides the location of sites cleaved by restriction enzymes.

Assume that one conducted a typical cloning experiment using a typical plasmid,transformed an appropriate host bacterial strain,and plated the bacteria on an appropriate X-gal medium.Blue and white colonies appeared.Which of the two types of colonies,blue or white,would most likely contain the recombinant plasmid?

In a typical PCR,primers are used to cleave specific regions of the DNA template.

List,in order,the steps usually followed in producing recombinant DNA molecules in a plasmid vector.

What is the specific application of reverse transcriptase in the preparation of cDNA?