C-terminal 80 Amino Acid Chemical Synthesis and Prokaryotic Expression of Swine Myostatin Gene

The coding sequence of swine myostatin optimized according to common codons of E.coli was divided 12 single segments,F 1,F 2,F 3,R 6,R 5,R 4,F 4,F 5,F 6,R 3,R 2 and R 1.These segments were synthesised by chemical means.Each pair of adjancent segments overlaps with 20 bp.The end of F 1 and R 1 are designed with recognition site sequence of restriction enzyme,NcoⅠ and XhoⅠ.A construction Mstnd pMD 18T was generated by inserting the sequence of 254 bp obtained by a PCR into pMD 18T vector and selecting the sense clones.The result showed that the cloned sequence coincides with the designed sequence by sequencing.This construction was digested with NcoⅠ and XhoⅠ and ligated the pET 28a vector digested with the same enzymes using T4 DNA ligase.And a construction was generated by transforming the plasmid MSTN pET 28a to the competent cell of BL21(DE3).The sense clone was induced with IPTG.The expression of swine myostatin gene was determined on SDS PAGE.