We have three plasmids. The first is the cloning vector, pGEM. The second is the

Question

We have three plasmids. The first is the cloning vector, pGEM. The second is the plasmid that would result if EcoRI had been used to construct the genomic library. The third is the plasmid that would result if BamHI had been used to construct the library. Fill out the table below with the expected number of DNA fragments that would result from digesting each plasmid with the restriction enzymes listed. Use figure 7 (attached).

Explanation / Answer

In pGEM , there's one site each for BamH1 & EcoR1. Thus in a simple pGEM molecule, BamH1 or EcoR1 alone would result in a single fragment, but when both the enzymes are used we will get 2 fragments. In pGEM +EcoR1 insert, number of sites for EcoR1 will increase by one. Thus, alone BamH1 will result in one fragment, EcoR1 will result into 2 fragments. If used together, we will have 3 fragments. Similarly, if pGEM+BamH1 insert is used, there will an increased site for BamH1. Plasmid BamH1 EcoR1 BamH1+EcoR1 pGEM 1 1 2 pGEM+ EcoR1 insert 1 2 3 pGEM +BamH1 insert 2 1 3

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