A.) A plasmid contains a multiple cloning site inside the coding region of the lacZ gene and also contains an ampicillin resistance gene at a separate locus. When cells are transformed with a successfully recombinant plasmid containing a piece of DNA cloned into the multiple cloning site, what color will the colonies be when grown on an agar plate containing ampicillin and X-gal?
(a.) green (b.) white (c.) colonies will not grow (d.) blue (e.) clear
B.) If one wishes to clone a gene using typical restriction endonucleases, how does the restriction endonuclease identify the appropriate cut sites in the genome?
(a.) the endonuclease cuts randomly in the genome
(b.) the endonuclease recognizes the gene of interest
(c.) the endonuclease identifies its specific recognition sequence
(d.) the endonuclease cannot identify the cut sites
C.) A scientist is troubleshooting the synthesis of a cDNA library. The scientist performs both a Northern and a Southern blot. The Northern blot demonstrated the presence of RNA while the Southern blot indicated that no cDNA was present in the sample. What is likely to be the cause of the failed synthesis of the cDNA library?
(a.) proper primers (b.) temperature too cold for annealing (c.) defective reverse transcriptase (d.) too many dNTPs
A.) A plasmid contains a multiple cloning site inside the coding region of the lacZ gene and also contains an ampicillin resistance gene at a separate locus. When cells are transformed with a successfully recombinant plasmid containing a piece of DNA cloned into the multiple cloning site, what color will the colonies be when grown on an agar plate containing ampicillin and X-gal?
(a.) green (b.) white (c.) colonies will not grow (d.) blue (e.) clear
B.) If one wishes to clone a gene using typical restriction endonucleases, how does the restriction endonuclease identify the appropriate cut sites in the genome?
(a.) the endonuclease cuts randomly in the genome
(b.) the endonuclease recognizes the gene of interest
(c.) the endonuclease identifies its specific recognition sequence
(d.) the endonuclease cannot identify the cut sites
C.) A scientist is troubleshooting the synthesis of a cDNA library. The scientist performs both a Northern and a Southern blot. The Northern blot demonstrated the presence of RNA while the Southern blot indicated that no cDNA was present in the sample. What is likely to be the cause of the failed synthesis of the cDNA library?
(a.) proper primers (b.) temperature too cold for annealing (c.) defective reverse transcriptase (d.) too many dNTPs
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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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In your
Which of the following did Erwin Chargaff observe?
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The melting temperature (Tm) of a DNA duplex is:
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Given your knowledge and the descriptions of each of thesemethods, which of the following does NOT rely on the ability ofnucleic acids to hybridize with each other?
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Which of the following is a possible reason why DNA uses thymineinstead of uracil?
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Which of the following is an enzyme used in cloning to breakcovalent bonds?
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Which of the following could you use to remove the 5' phosphatesafter cleavage of a plasmid with a single type of restrictionenzyme to ensure that the plasmid would not simply fuse backtogether upon addition of ligase?
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Which of the following would you use to create a cDNA librarythat you would not use for a genomic library?
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A plasmid vector typically has which of the followingfeatures?
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Which of the following is FALSE of genomic or cDNAlibraries?
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Which of the following is NOT a typical component of apolymerase chain reaction (PCR)?
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How can a fusion protein be formed?
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Why is green fluorescent protein (GFP) so useful in visualizingfusion proteins in eukaryotes?
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You wish to determine the cellular location of a protein butunfortunately it is not particularly antigenic (can t develop astrong antibody response). How could you solve this problem?
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A positive result for the yeast two-hybrid analysis means thefollowing:
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Which of the following is the approximate size of the humangenome?
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Which of the following is correct about the structure orlocation of genes?
Question 22 options:
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