Exam 20: Recombinant DNA Technology

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

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

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

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

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.

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

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

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.

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

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

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

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

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

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

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.

E.coli is a common YAC.

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

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?

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