Bacterial cells are used to express proteins from other organisms. Your lab has isolated a human gene that encodes an enzyme that want to produce in a bacterial cell. You can stably transfer the human gene, along with its promoter into a bacterial cell but no human protein is made. Assume that youâve done the procedure correctly and have established that the sequence youâve used is the reason for the lack of protein. How can this be explained? 1.5 pts
Once you solved this initial problem, you can detect RNA from the human gene in the bacterial culture. However, the mRNA is twice as long as the mRNA isolated from this gene in humans and the enzyme coded for by the gene does not appear to be functional. Explain why. 1.5 pts
Bacterial cells are used to express proteins from other organisms. Your lab has isolated a human gene that encodes an enzyme that want to produce in a bacterial cell. You can stably transfer the human gene, along with its promoter into a bacterial cell but no human protein is made. Assume that youâve done the procedure correctly and have established that the sequence youâve used is the reason for the lack of protein. How can this be explained? 1.5 pts
Once you solved this initial problem, you can detect RNA from the human gene in the bacterial culture. However, the mRNA is twice as long as the mRNA isolated from this gene in humans and the enzyme coded for by the gene does not appear to be functional. Explain why. 1.5 pts
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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.)
Skip to question text.
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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Skip to question text.
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 |
Flag this Question
In your
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.)
Skip to question text.
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 |
Flag this Question
Plasmids (or vectors) are important in biotechnology becausethey are
| a vehicle for the insertion ofrecombinant DNA into bacteria. |
| surfaces for respiratoryprocesses in bacteria. |
| recognition sites onrecombinant DNA strands. |
| surfaces for protein synthesisin eukaryotic recombinants. |
| proviruses incorporated intothe host DNA |
Flag this Question
Plasmids are put into bacterial cells by
| restriction enzymes |
| DNA ligase |
| binding of cohesive stickyends |
| transformation |
Flag this Question
Restriction enzymes usually
| cut donor DNA evenly so smoothedges result |
| cut donor DNA but do notaffect plasmids |
| make staggered cuts atspecific sequences in DNA in both donor DNA and plasmid |
| are used in ligating plasmidsinto bacterial host cells |
| more than one of theabove |
Flag this Question
After combining DNA fragments in a cloning experiment, ___ isused to covalently join the DNA segments.
| Restriction enzyme |
| DNA Ligase |
| Reverse transcriptase |
| DNA polymerase |
| Helicase |
Flag this Question
It is theoretically possible for a gene from any organism tofunction in any other organism. Why is this possible?
| All organisms haveribosomes. |
| All organisms have the samegenetic code. |
| All organisms are made up ofcells. |
| All organisms have similarnuclei. |
| All organisms have transferRNA. |
Flag this Question
Skip to question text.
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 withrestriction enzyme X and insert the fragments cut with Y into theplasmid. |
| cut the plasmid with enzyme Xand then insert the gene into the plasmid. |
| cut the DNA again withrestriction enzyme Y and insert these fragments into the plasmidcut with the same enzyme. |
| cut the plasmid twice withrestriction enzyme Y and ligate the two fragments onto the ends ofthe human DNA fragments cut with restriction enzyme X. |
| insert the fragments cut withX directly into the plasmid without cutting the plasmid. |
Flag this Question
Which of the following is/are false in regard to expressionplasmids (also called expression vectors)?
| They are used to make proteinsusing a cloned gene. |
| They contain a promotor. |
| They are the first plasmidtype used to clone a gene. |
| They contain aterminator. |
| More than one of the above isfalse. |
Flag this Question
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 genecontains introns the bacteria will not remove them and theresulting amino acid sequence will be different that that made by aeukaryote. |
| The bacteria may not fold theprotein correctly. |
| The bacteria may degrade theprotein. |
| All of the above are potentialproblems. |
Flag this Question
Cloning allows for production of proteins in much larger amountsthan occurs in the cells from which the gene is isolated.
| True |
| False |
Flag this Question
Question 111 pts
Gene cloning is used to do all of the following except
| Make insulin |
| Making genetically identicalanimals (e.g. Dolly the sheep) |
| Make vaccines |
| Perform Gene Therapy |
| Making genetically engineeredplants |
Flag this Question
In your
