You extract DNA from some of your cells and cut out the complete human growth hormone gene directly from this DNA. You put this gene into a plasmid and transform E.coli with the plasmid. To your dismay, you find that E. coli does not make any human growth hormone. What is the most likely explanation for the failure of this experiment?
A) Bacteria cannot carry out RNA splicing to remove introns and so produced a much larger protein.
B) Bacteria lack a nucleus for proper transcription of eukaryotic genes.
C) Human DNA cannot be cloned in a bacterium.
D) Human DNA can be maintained in cloned form only for brief periods in bacteria.
You extract DNA from some of your cells and cut out the complete human growth hormone gene directly from this DNA. You put this gene into a plasmid and transform E.coli with the plasmid. To your dismay, you find that E. coli does not make any human growth hormone. What is the most likely explanation for the failure of this experiment?
A) Bacteria cannot carry out RNA splicing to remove introns and so produced a much larger protein.
B) Bacteria lack a nucleus for proper transcription of eukaryotic genes.
C) Human DNA cannot be cloned in a bacterium.
D) Human DNA can be maintained in cloned form only for brief periods in bacteria.
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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 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. |
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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 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. |
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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 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
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 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 |
Flag this Question
Plasmids are put into bacterial cells by
restriction enzymes |
DNA ligase |
binding of cohesive sticky ends |
transformation |
Flag this Question
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 |
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 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. |
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 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. |
Flag this Question
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. |
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 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. |
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 identical animals (e.g. Dolly thesheep) |
Make vaccines |
Perform Gene Therapy |
Making genetically engineered plants |
Flag this Question
In your
You decide to identify the CFTR mutation by analyzing the genomic DNA of your patients compared to healthy individuals. You specifically are looking to see whether a specific 3' gene truncation has occurred in the patients. You will determine this using hybridization techniques with samples from healthy and CF patients. Which of the following will allow you to accomplish this?
Using an RNA probe complementary to the region not removed by the truncation. | |
Using an RNA probe complementary to the region removed by the truncation. | |
Using an DNA probe complementary to the region not removed by the truncation. | |
Using an DNA probe complementary to the region removed by the truncation. |
To conduct the hybridization experiment, you are trying to decide between using a DNA or RNA probe. Which would be ideal to use and why?
As both are composed of nucleic acids, using either would result in identical results. | |
An RNA probe because RNA has uracil bases. | |
An RNA probe because it could also be used in a translation experiment. | |
A DNA probe because it is more stable than RNA. | |
A DNA probe because RNA cannot bind to DNA. |
Imagine Hershey/Chase had used an RNA virus (genome composed of RNA) instead of a DNA virus in their experiment. Would radioactivity still have been found in the pellet?
No, because only DNA can be labeled with radioactivity. | |
No, because the RNA genome would not enter the bacteria upon infection. | |
No, because while DNA and RNA nucleotides are similar, they are not identical. | |
Yes, because DNA and RNA nucleotides are similar. | |
Yes, because genome in any form (DNA, RNA, protein) would be labeled similarly. |
The human genome consists mostly of non-coding DNA. Which of the following are benefits of this?
Random DNA mutations generally won't affect RNA and protein function. | |
It is faster to duplicate the genome when these are present. | |
The existence of introns can lead to multiple variations of proteins encoded by a single gene. | |
It is unlikely transposons would exist in the genome if there was too much protein coding DNA. |
You accidentally add a mutant dNTP (which has an H instead of an OH connected to the 3â carbon) to a reaction where DNA is being replicated. Which of the following is true of this mutant dNTP?
It can be incorporated into DNA strand but cannot form a phosphodiester bond with an incoming wild type dNTP | |
It can be incorporated into a DNA strand but cannot base pair with a complementary nucleotide | |
It can be incorporated into a DNA strand and can form a phosphodiester bond with an incoming dNTP, but only if it is another mutant dNTP | |
It cannot be incorporated into a DNA strand. |
Andrew Murray's sister, Andrea, is adding to her brother's work on chromosomes. She is using cells that are unable to synthesize adenine (âade) and histidine (âhis). The plasmid she is currently working with consists of an origin of replication and the Ade gene.
Following her transformation of the plasmid into her yeast, what media will the cells be plated on to select for cells that have picked up the plasmid?
Media containing histidine | |
Media containing adenine | |
Media lacking adenine | |
Media lacking histidine |
Andrew Murray's sister, Andrea, is adding to her brother's work on chromosomes. She is using cells that are unable to synthesize adenine (âade) and histidine (âhis). The plasmid she is currently working with consists of an origin of replication and the Ade gene.
She starts by attempting to add the centromere DNA into a plasmid containing the origin of replication. Unfortunately, when adding the centromere sequence, the origin of replication is removed, thus leaving a plasmid with only a centromere and selection marker. Following plasmid transformation, what growth result will she see on her plates?
No colonies | |
Little colonies | |
Big colonies |
Andrew Murray's sister, Andrea, is adding to her brother's work on chromosomes. She is using cells that are unable to synthesize adenine (âade) and histidine (âhis). The plasmid she is currently working with consists of an origin of replication and the Ade gene.
She fixes the mistakes from the previous experiment and now has a complete plasmid (selection marker, origin of replication, centromere). She then inserts telomere sequences into the plasmid. How will this impact her transformation?
It will not impact her transformation | |
Her transformation will no longer work because plasmids donât require telomeres | |
She will now see much larger colonies | |
She will now see fewer but larger colonies | |
She will now see smaller colonies |
In the Meselson/Stahl experiment, E. coli were first grown in media containing heavy nitrogen, 15N, and then transferred to light nitrogen, 14N, at the beginning of the experiment.
Imagine that their data showed that replication occurs in a conservative manner instead of semi-conservative. What fraction of the DNA helices will consist of mixed DNA after 4 rounds of replication in this case?
None | |
More than 75% | |
25-50% | |
51-75% | |
Less than 25% |