Trichoderma Cellobiohydrolase CBHâ…¡ Gene Cloning and Expression in Escherichia Coli

Abstract

In this study, cbhâ…¡ (cellobiohydrolase) gene from Trichoderma viride. The reference to GenBank published viride (T. reesei) cbhâ…¡ gene (GenBank: M55080) obtain cellobiohydrolase gene cbhâ…¡ of cDNA sequence 1441bp, after codon optimization, sent synthesized by Sangon Biotech (Shanghai, China). The target gene shuttle vector pMG36e connection with prokaryotic expression vector pMG36e-CBHâ…¡; The use of heat shock method which transformed into E. coli BL21 competent cells to construct the recombinant bacteria, fungi and culturing the recombinant proteins were extracted, by SDS-PAGE electrophoresis, to obtain a target protein of approximately 53kD band; Measured by DNS method bacterial cell lysate supernatant after sonication, the activity maximum 16.4U / mL, cell lysate precipitated almost no activity, the results show cellobiohydrolase genes in E. coli successfully was expressed.