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Inserting the vgp gene into a cloning vector (chapter 20)
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Part B - Inserting the vgp gene into a cloning vector
In the second step of the cloning procedure, DNA fragments produced by a restriction enzyme are inserted into a cloning vector that has been cut by the same enzyme. The vector used in this tutorial is a plasmid.
To clone the vgp gene into the plasmid below, a restriction enzyme must do all of the following:
cut the human DNA on both sides of the vgp gene, but not inside the gene
cut the plasmid without cutting inside the ampicillin resistance gene (ampR ), shown in blue (The importance of the ampR gene will be explained in Part C.)
produce sticky ends when it cuts both the human DNA and the plasmid, so that the human DNA fragments can combine with the plasmid

The questions below will help determine which restriction enzyme should be used to make a plasmid that contains both an intact vgp gene and a functional ampR gene.
Answer questions 1–4 by selecting from the answer choices on the left. Drag the correct answer to the right of each question.


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Inserting the vgp gene into a cloning vector (chapter 20)
Jun 15, 2013

a) BamHI - Yes, produces sticky ends, and cuts around the gene
b) HindIII - Yes, produces sticky ends, and cuts around the gene
c) EcoRI - No, produces sticky ends, but cuts IN the gene
d) HaeIII - No, produces blunt ends, but does cut around the gene



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Inserting the vgp gene into a cloning vector (chapter 20)
Jun 15, 2013

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This is the correct Answer:
1)BamHI , Haelll and HindIIl
2) BamHI, EcoRI, and HaeIII,
3) BamHI, EcoRI, and HindIII,
4) BamHI only.

BamHI cuts the plasmid outside the ampR gene, leaving the gene intact and functional, which is important for the next step in the cloning procedure. Once BamHI opens the plasmid, the sticky ends it produces in the plasmid can base-pair with those on BamHI human DNA fragments. If DNA ligase is then added, it will covalently join the sugar-phosphate backbones of the plasmid and the fragments, producing recombinant plasmids.

Note that human DNA contains many BamHI restriction sites in addition to the two shown in Part A. Therefore, only a small fraction of the recombinant plasmids will have the vgp gene, like the plasmid shown above. Most of the recombinant plasmids will have a fragment of human DNA that does not contain the vgpgene, and would not be of interest to the scientists.




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Inserting the vgp gene into a cloning vector (chapter 20)
Jun 15, 2013

1)BamHI , Haelll and HindIIl
2) BamHI, EcoRI, and HaeIII,
3) BamHI, EcoRI, and HindIII,
4) BamHI only.

Just finish my home work You can trust this answer Kiss



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Inserting the vgp gene into a cloning vector (chapter 20)
Jun 15, 2013

Thank you so much ochorios and our guest George both of you were correct.. both of you are life savers..Bio has been killing me Sad



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Jun 15, 2013

noooo problem Smug



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Nov 9, 2013

Excellent, thank you!!!



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Nov 17, 2013

Thanks for the help guys and the explanation really clarified everything



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Nov 17, 2013

Thank you ♥



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Nov 20, 2013

thank you!!



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Nov 20, 2013

Thanks!



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Nov 24, 2013

THANKYOU SO MUCH!



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Dec 1, 2013

You are appreciated!



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Dec 1, 2013

Thanks for posting this !!!



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