PLANTBI C112 Study Guide - Midterm Guide: Reca, Plasmid, Electroporation

1.Provide one example of a sequence-specific DNA binding protein, and explain its function. Also provide one example of a non-squence specific DNA binding protien, and explain its function.

2.. Specialized plasmid vectors can be used to easily identify E. coli celss containing recombinant plasmids (as opposed to E. coli cells containing vector self-ligations). Describe specifically how one of these systems works.

View/perform/read ALL THREE of the following prior to answeringthe questions.

http://highered.mcgraw-hill.com/olcweb/cgi/pluginpop.cgi?it=swf::535::535::/sites/dl/free/0072437316/120078/micro10.swf::Stepsin Cloning a Gene (Links to an external site.)

http://www.discoverbiotech.com/wiki/-/wiki/Main/Applications ofCloning (Links to an external site.)

http://www.wiley.com/college/boyer/0470003790/animations/cloning/cloning.htm(Links to an external site.)

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From the list below, which of the following is the most logicalsequence of steps for splicing foreign DNA into a plasmid andinserting the plasmid into a bacterium?
I. Transformbacteria with recombinant DNA molecule
II. Cutthe plasmid DNA using restriction enzymes
III. Extractplasmid DNA from bacterial cells
IV. Hydrogen-bondthe plasmid DNA to nonplasmid DNA fragments
V. Useligase to seal plasmid DNA to nonplasmid DNA

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Plasmids (or vectors) are important in biotechnology becausethey are

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Plasmids are put into bacterial cells by

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Restriction enzymes usually

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After combining DNA fragments in a cloning experiment, ___ isused to covalently join the DNA segments.

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It is theoretically possible for a gene from any organism tofunction in any other organism. Why is this possible?

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Assume that you are trying to insert a gene into a plasmid andsomeone gives you a DNA sample cut with restriction enzyme X. Thegene you wish to insert from the given sample has sites on bothends for cutting by restriction enzyme Y. You have a plasmid with asingle site for Y, but not for X. Your strategy should be to

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Which of the following is/are false in regard to expressionplasmids (also called expression vectors)?

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What is NOT a potential problem(s) associated with usingbacteria containing a cloned eukaryotic gene (e.g. a human gene) toproduce a functional protein?

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Cloning allows for production of proteins in much larger amountsthan occurs in the cells from which the gene is isolated.

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Question 111 pts

Gene cloning is used to do all of the following except

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In your

A-Which of the following is NOT one of the steps required toclone a piece of DNA using a plasmid vector?
1. Cut a vector and a DNA insert with aspecific restriction enzyme to create complementary "stickyends."
2. Isolate DNA fragments by gelelectrophoresis.
3. Ligate insert DNA and a vector withsimilar sticky ends together.
4. Transform ligated pieces of DNA into E.coli.
5. Select the plasmid vector that hasreplaced its origin of replication with insert DNA.

B-A representative genomic library of anorganism's DNA fragments results from the use of
1. DNA ligase joining multiple types ofvectors.
2. partial digestion with frequentlycutting restriction endonucleases.
3. many pieces of recombinant DNA fromseveral different types of organisms.
4. viruses that cleave the DNA into randompieces.

C-One of the most usefulmethods for identifying a specific DNA sequence is
1. the Southern blot.
2. the Western blot.
3. magnetic resonance imaging.
4. the Eastern blot.
5. thin-layerchromatography.