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| Optimizing the Soluble Expression Conditions of Sindbis Virus E2 Envelope Protein |
| LI Jiang-jiao, ZHU Wu-yang, HE Ying, LIANG Guo-dong |
| State Key Laboratory for Infectious Disease Prevention and Control, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China |
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Abstract To improve the soluble expression level of extracellular domain of E2 envelope protein of Sindbis Virus (SINV) in prokaryotic expression system, the expression conditions were optimized. The extracellular domain of E2 gene was cloned from cDNA of SINV and sub cloned into expression vector pGEX-6P-1. The recombinant plasmid pGEX-6p-1-E2 was constructed and transformed into four host bacteria respectively. The optimum host bacteria and induction temperature were selected by SDS-PAGE assay. The molecular chaperone co-expression system was constructed and the optimum molecular chaperone vector was screened. The results were described as follows: the E2 gene was amplified and cloned into vector pGEX-6p-1 successfully; the quantity of E2 expression was similar in four host bacteria; the soluble expression level of E2 was highest when induced at 16℃; in the molecular chaperone co-expression system, the five vectors containing different molecular chaperones (pG-KJE8、 pGro7、pKJE7、 pG-Tf2 and pTf16) improved the soluble expression level of E2 in varying degrees when co-expression with recombinant plasmid pGEX-6p-1-E2, and pG-Tf2 was confirmed to contain the optimum molecular chaperones combination and made the soluble expression level of E2 improved 15.7%. These results can provide a potentia1 value for further structural and functional studies of E2.
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Received: 31 August 2010
Published: 18 February 2011
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