细菌素Bacteriocin E50-52(H)基因设计及其毕赤酵母表达载体构建

Gene Design of Bacteriocin E50-52 and Construction of its Pichia pastoris Expression Vector

设计及合成细菌素Bacteriocin E50-52(H)基因,克隆到组成型分泌表达质粒pGAPZαA中构建重组质粒pGAPZα-Bacteriocin E(H),经PCR、测序验证正确后,电转化整合到毕赤酵母基因组,基因组PCR、测序验证结果表明成功构建了细菌素组成型重组表达载体,为在毕赤酵母中表达奠定了基础。

Bacteriocin E50-52 gene was designed according to its amino acid sequence and cloning site of constitutive secreted plasmid pGAPZαA,then synthesized,inserted into pMD18-T.Recombinant pGAPZα-Bacteriocin E was constructed by ligating the digested target gene and pGAPZαA with XhoⅠ and XbaⅠ.Recombinant Pichia pastoris was constructed by transforming linearized recombinant plasmid in strain SMD1168.Sequencing result of recombinant vector and genome of Pichia pastoris indicated this expression plasmid was successfully constructed.