Construction, expression and purification of recombinant somatostatin
AIM: To clone and express pig somatostatin gene in Escherichia. Coli (E. coli) and to purify and identify the fusion protein. METHODS: The gene of pig somatostatin was designed and synthesized according to GenBank data and optimized to be consistent with preferred codens of E, coli. Then the synthesized SS gene was cloned into pGEX-4T-1 prokaryotic expression vector and the plasmid was transformed into E. coli strain DH5α. GST-SS fusion protein was expressed with IPTG induction and purified by GST-affinity chromatography. RESULTS: Restriction enzyme digestion and DNA sequencing showed that the sequence of SS gene was correct. The GST-SS fusion protein was expressed at high level with IPTG induction. After the E. coli was lysed by ultrasonic wave and lysozyme, the recombinant protein was purified with Glutathione Sepharose 4B affinity chromatography. CONCLUSION: The fusion protein GST-SS is effectively expressed in E. coli and the purity is over 95% after affinity chromatography, which lays a good foundation for large-scale production and optimization of the recombinant vaccine for wide application in animal husbandry.
NETL
NSTL
万方数据
维普
