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Concepts of Genetics

Biology

Quiz 20 :

Recombinant Dna Technology

Quiz 20 :

Recombinant Dna Technology

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

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What is meant by the designation EcoRI?
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one of the first restriction enzymes isolated from E. coli

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List, in order, the steps usually followed in producing recombinant DNA molecules in a plasmid vector.
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isolation of DNA (foreign and plasmid), digestion of DNAs with an appropriate restriction endonuclease, ligation of fragments, transformation of host cells

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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?
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Nucleic acid blotting is widely used in recombinant DNA technology. In a Southern blot, one generally ________.
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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.
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List two especially useful characteristics of cloning vectors.
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In which of the following biochemical reactions is it common to use ddNTPs (dideoxyribonucleoside triphosphates)?
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What might be a reasonable function of restriction endonucleases in a bacterium, distinct from their use by molecular biologists?
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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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Some vectors such as pUC18 and others of the pUC series contain a large number of restriction enzyme sites clustered in one region. Which term is given to this advantageous arrangement of restriction sites?
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In the context of molecular genetics, reverse transcription PCR (RT-PCR) refers to ________.
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In the context of recombinant DNA technology, what is meant by the term vector?
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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.
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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?
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A ddNTP, used often in DNA sequencing, lacks a(n) ________ at the ________ and ________ carbons.
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Molecular biologists rely on many, often sophisticated, techniques to pursue their discipline. One may list ultracentrifugation, electron microscopy, X-ray diffraction, electrophoresis, and computer interfacing as fundamental tools. Model organisms provide the raw materials for study. List three "organisms" (or organismic groups) often used by recombinant DNA technologists and describe a major advantage of each group.
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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.
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One of the primary reasons for generating a large number of clones in a eukaryotic genomic library is that ________.
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What is recombinant DNA technology? What would you list as safety issues associated with recombinant DNA technology?
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