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Social Science16. 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 bacteriaB: is a kind of product that DNA ligase cannot formC: makes E. coli sensitive to antibioticsD: lacks a replication originE: is beneficial to bacteriaom
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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
Expert Answer
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.
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).
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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