Question
Asked Nov 3, 2019
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16. Why is re-circularized insert generally not a problem in cloning procedures, while re-
circularized vector can be a problem? Is it because re-circularized insert:
A: is detrimental to bacteria
B: is a kind of product that DNA ligase cannot form
C: makes E. coli sensitive to antibiotics
D: lacks a replication origin
E: is beneficial to bacteria
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16. Why is re-circularized insert generally not a problem in cloning procedures, while re- circularized vector can be a problem? Is it because re-circularized insert: A: is detrimental to bacteria B: is a kind of product that DNA ligase cannot form C: makes E. coli sensitive to antibiotics D: lacks a replication origin E: is beneficial to bacteria om

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Expert Answer

Step 1

Cloning or DNA cloning is the process of making multiple identical copies of the gene of interest by inserting it into a vector or plasmid. The insert (gene segment of interest) gets replicated each time the bacterium containing cloned plasmid, replicates.

Step 2

Steps of cloning includes:

  • Digesting and ligating the plasmid and gene of interest with the help of DNA restriction enzyme and DNA ligase.
  • Insertion of recombinant plasmid in the bacteria and selection of bacteria that took up the desired construct using selection markers (antibiotics).
  • Growing bacteria that took up the desired construct and harvesting the desired protein (encoded by the gene of interest).
Step 3

During the ligation of insert and the vector, sometimes the insert and vector re-circularizes with itself. The re-circularization of insert i...

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