Question

If purP gets incorporated into pCM999, it might be inserted in the “forward” orientation or the...

If purP gets incorporated into pCM999, it might be inserted in the “forward” orientation or the “reverse” orientation. Is one orientation preferable to the other? If so, how can you tell which plasmids have purP in the correct orientation? (Hint: do a restriction digest and run a gel. Which enzyme or enzymes will you use? What fragment sizes will you expect?)

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Answer #1

Length of pupR is 1400 bp

Lenght of pCM999 is 4000 bp.

AS we can see from the figure that during forward orientation we will two bands one of 2000 bp and another one of 3400 bp when digested with NotI and KpnI(KpnI is present in the gene and NotI in the vector 0 whereas in case of reverse orientation we will two bands one of 700 bp and another one of 4700 bp

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