Expression of carboxylesterase gene LmCarE25 from Locusta migratoria

Author of the article:WANG Jian-Xin

Author's Workplace：Institute of Applied Biology, Shanxi University

Key Words： Locusta migratoria, Carboxylesterase, Real-time PCR, Prokaryotic expression, Eukaryotic expression

Abstract：Carboxylesterases (CarE) are a family of multifunctional enzymes widely distributed among organism, and distinctly related to insecticide resistance. In this present study the physiological functions of the carboxylesterase member LmCarE25 was investigated from Locusta migratoria(Meyen). Real-time PCR rresults showed that the LmCarE25 was expressed in all of the adult tissues, while had lower expression levels in gastric cecum, wings and muscles, and higher expression levels in midgut, Malpighian tubules and fat body. In addition, colibacillus and insect cell expression system were used to express this enzyme in vitro. The recombinant LmCarE25 was successfully expressed by recombinant vector pET32a-LmCarE25 in E. coli BL21(DE3) and JM109, and its molecular weight was approximately 60 kDa, although it was inclusion bodies. Then LmCarE25 was subcloned into eukaryotic expression vector pFastBacHTA, the recombinant bacmid DNA was obtained by Bac-to-Bac® system, and was transfected into sf9 cells by lipid mediated transfection. Western-blot results suggested LmCarE25 expressed in soluble form at low level. These results might provide information for further study the function of carboxylesterase family of Locusta migratoria.