1. You have cloned a human gene along with its promoter and ribosome-binding site in a bacterial cloning vector and then introduced the recombinant plasmid into a bacterial host cell that supports the replication and segregation of the cloning vector. Unfortunately, even though you know the whole human gene sequence is present (by DNA hybridization); you cannot get expression of the gene. Which of the following is not a reason why there is no expression?
A. The vector cannot replicate
B. The human promoter is not recognized by the bacterial host cell
C. The ribosome-binding site is not recognized by the bacterial host cell
D. The human gene lacks introns, so it is not transcribed by the host bacterium
1. You have cloned a human gene along with its promoter and ribosome-binding site in a bacterial cloning vector and then introduced the recombinant plasmid into a bacterial host cell that supports the replication and segregation of the cloning vector. Unfortunately, even though you know the whole human gene sequence is present (by DNA hybridization); you cannot get expression of the gene. Which of the following is not a reason why there is no expression?
A. The vector cannot replicate
B. The human promoter is not recognized by the bacterial host cell
C. The ribosome-binding site is not recognized by the bacterial host cell
D. The human gene lacks introns, so it is not transcribed by the host bacterium
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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
| IV, V, I, II, III |
| III, II, IV, V, I |
| III, IV, V, I, II |
| II, III, V, IV, I |
| I, II, IV, III, V |
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Plasmids (or vectors) are important in biotechnology becausethey are
| a vehicle for the insertion of recombinant DNA intobacteria. |
| surfaces for respiratory processes in bacteria. |
| recognition sites on recombinant DNA strands. |
| surfaces for protein synthesis in eukaryotic recombinants. |
| proviruses incorporated into the host DNA |
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Plasmids are put into bacterial cells by
| restriction enzymes |
| DNA ligase |
| binding of cohesive sticky ends |
| transformation |
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Restriction enzymes usually
| cut donor DNA evenly so smooth edges result |
| cut donor DNA but do not affect plasmids |
| make staggered cuts at specific sequences in DNA in both donorDNA and plasmid |
| are used in ligating plasmids into bacterial host cells |
| more than one of the above |
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After combining DNA fragments in a cloning experiment, ___ isused to covalently join the DNA segments.
| Restriction enzyme |
| DNA Ligase |
| Reverse transcriptase |
| DNA polymerase |
| Helicase |
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It is theoretically possible for a gene from any organism tofunction in any other organism. Why is this possible?
| All organisms have ribosomes. |
| All organisms have the same genetic code. |
| All organisms are made up of cells. |
| All organisms have similar nuclei. |
| All organisms have transfer RNA. |
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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
| cut the plasmid with restriction enzyme X and insert thefragments cut with Y into the plasmid. |
| cut the plasmid with enzyme X and then insert the gene into theplasmid. |
| cut the DNA again with restriction enzyme Y and insert thesefragments into the plasmid cut with the same enzyme. |
| cut the plasmid twice with restriction enzyme Y and ligate thetwo fragments onto the ends of the human DNA fragments cut withrestriction enzyme X. |
| insert the fragments cut with X directly into the plasmidwithout cutting the plasmid. |
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Which of the following is/are false in regard to expressionplasmids (also called expression vectors)?
| They are used to make proteins using a cloned gene. |
| They contain a promotor. |
| They are the first plasmid type used to clone a gene. |
| They contain a terminator. |
| More than one of the above is false. |
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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?
| If the eukaryotic gene contains introns the bacteria will notremove them and the resulting amino acid sequence will be differentthat that made by a eukaryote. |
| The bacteria may not fold the protein correctly. |
| The bacteria may degrade the protein. |
| All of the above are potential problems. |
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Cloning allows for production of proteins in much larger amountsthan occurs in the cells from which the gene is isolated.
| True |
| False |
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Question 111 pts
Gene cloning is used to do all of the following except
| Make insulin |
| Making genetically identical animals (e.g. Dolly thesheep) |
| Make vaccines |
| Perform Gene Therapy |
| Making genetically engineered plants |
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