Cloning of Porcine Follistatin Gene Mature Peptide Sequence and Expression in E.coli BL21(DE3)

A pair of primers was designed according to the porcine follistatin(FS)gene mature peptide sequence(GenBank:M36512.1),the mature peptide sequence of FS gene was achieved by PCR.The PCR product of the expected length was cloned into the pMD18-T vector and transformed into E.coli DH5α.The positive clones were identified by PCR,enzyme digestion and sequencing.Then insert the FSgene mature peptide between the BamHⅠand HindⅢsites of the pRSET-A vector generating the recombinant plasmid pR-FS and transformed it into E.coli BL21(DE3).The positive clones were also identified by PCR,enzyme digestion and sequencing.Bacteria transformed by pR-FS were expressed in E.coli BL21(DE3)after induction with IPTG and produced a product showing expected molecular mass of 26.1ku in SDS-PAGE,which achieved the highest level induced by 0.2mmol/L IPTG for 3h.The product can be purified by 50% Ni-NTA agarose.These results demonstrated that the porcine FSgene mature peptide sequence was successfully cloned and expressed in E.coli BL21(DE3).