A student wanted to clone a gene G (size–1.6 kb) in a vector V (size - 5 kb). The vector has sites for restriction enzymes RE1, RE2, and RE3 in its multiple cloning site. The site for RE2 is located in between the sites for RE1 and RE3.

A student wanted to clone a gene G (size&ndash;1.6 kb) in a vector V (size - 5 kb). The vector has sites for restriction enzymes RE1, RE2, and RE3 in its multiple cloning site. The site for RE2 is located in between the sites for RE1 and RE3.
The student carried out the following steps for cloning:
Step 1. Cut the gene G with restriction enzymes RE1 and RE3
Step 2. Cut the vector V with restriction enzymes RE1 and RE3. Step
3. Ligated G and V from Steps 1 and 2, transformed into E.coli and isolated plasmed L from the transformants
The plasmid L was cut with RE2, which gave a band of size-6.6 kb.
The following three reasons were given to explain this observation:
P. Plasmid L is same as the vector V, since E. coli got transformed only with uncut V.
Q. Gene G has a site for RE2.
R. RE1 and RE2 are isoschizomers.
Which of the above three reasons are possible?

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M.Sc. Entrance

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