Exam 20: Recombinant DNA Technology

Over the years,sophisticated plasmid vectors have been developed for use in recombinant DNA technology.List at least two features that have been introduced in particularly useful vectors.

Assume that a plasmid (circular)is 3200 base pairs in length and has restriction sites at the following locations: 400,700,1400,2600.Give the expected sizes of the restriction fragments following complete digestion.

Words such as level,civic,and kayak have something in common compared to the fundamental tool of recombinant DNA technology.In the context of recombinant DNA technology,which term would be used to describe such words?

The function of a ddNTP in DNA sequencing is to methylate guanine.

Some restriction enzymes cleave DNA in such a manner as to produce blunt ends.Most often ligation of blunt end fragments is enhanced by the use of the enzyme terminal deoxynucleotidyl transferase.Speculate on the function of deoxynucleotidyl transferase in terms of using blunt end fragments in cloning.

What is recombinant DNA technology? What would you list as safety issues associated with recombinant DNA technology?

A ddNTP,used often in DNA sequencing,lacks a(n)________ at the ________ and ________ carbons.

In what way are specific DNA sequences of the template amplified in the polymerase chain reaction? In other words,how does one target the target?

Under ideal conditions,how many copies of all the sequences of the host genome should be represented in a genomic library?

The PCR (polymerase chain reaction)protocol that is currently used in laboratories was facilitated by the discovery of a bacterium called Thermus aquaticus in a hot spring inside Yellowstone National Park,in Wyoming.This organism contains a heat-stable form of DNA polymerase known as Taq polymerase,which continues to function even after it has been heated to 95ᵒC.Why would such a heat-stable polymerase be beneficial in PCR?

In which of the following biochemical reactions is it common to use ddNTPs (dideoxyribonucleoside triphosphates)?

In a PCR,primers are complementary to stretches of DNA with which they anneal.

What method might be used to study a knockout mouse that,by virtue of the lost gene or genes,generates a lethal condition?

Assume that you have cut λ DNA with the restriction enzyme HindIII.You separate the fragments on an agarose gel and stain the DNA with ethidium bromide.You notice that the intensity of the stain is less in the bands that have migrated closer to the "+" pole.Give an explanation for this finding.

One of the primary reasons for generating a large number of clones in a eukaryotic genomic library is that ________.

A common term for a plasmid or other DNA element that serves as a cloning vehicle is vector.

Of what advantage is it to have a multiple cloning site (multiple unique restriction sites)embedded in the lacZ component of a plasmid?

During a PCR,heat is provided to inactivate the polymerase enzyme.

In the context of recombinant DNA technology,what is meant by the term vector?

What is the name of the process by which bacterial colonies (cells)are transferred from one agar plate to another,maintaining the same spatial pattern?