For
directional cloning, a BamHI site was introduced to the ends of the PCR products. A BamHI
site was also located on the multiple cloning site of pQE-T. The PCR products were ligated
with pQE-T. The directionally inserted recombinants were distinguished by using BamHI
to digest the recombinants because there are two BamHI sites located on the both sides
of PCR fragment. In order to identify the T-vector functions, the 14-3-3-ZsGreen and hRBP
genes were amplified and a BamHI site was added to the ends of the genes to confirm this
vector by ligation with pQE-T. Results showed that the 14-3-3-ZsGreen and hRBP were cloned
to the vector pQE-T directly and corresponding proteins were successfully produced