The restriction enzyme HincII was isolated from the bacterium Haemophilas influe

Question

The restriction enzyme HincII was isolated from the bacterium Haemophilas influenza. The DNA sequence cut by this enzyme is shown here. (Note that Y means any pyrimidine--C or T--and that R means any purine--A or G.) CTY*RAC GAR*YTG HincII is a blunt-end cutter that cuts after the Y. ou cut a gene of interest with HincII and would like to insert it into the multiple cloning site (MCS) of a plasmid. The plasmid's MCS contains the cut sites listed below. Which of these sites could you use to insert the gene of interest? Select all that apply.

A. GTC*AAC

CTG*TTG

B. GGT*ACC

CCG*TGG

C. GTA*TAC

CAT*ATC

D. GT*CAAC

CAGT*TG

E. G*TYRAC

CGRYT*G

F. none of these sites

Explanation / Answer

The enzyme HindII recognizes the following sequence:

CTY*RAC

GAR*YTG

Where Y can be either C or T, and R can be either G or A. As the enzyme is a blunt-end cutter, the only cloning sites in which the gene can be cloned, also need to have a blunt end

Therefore the answers are:

A. GTC*AAC

CTG*TTG

B. GGT*ACC

CCG*TGG

C. GTA*TAC

CAT*ATC

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