中文摘要：

[目的]建立金环蛇抗菌肽Cath-BF的毕赤酵母高效分泌表达系统。[方法]Cath-BF基因用引物重叠延伸法合成,克隆于p GAPZa-A,电转化毕赤酵母SMD1168,博莱霉素筛选高抗性转化子,以痤疮丙酸杆菌筛选抗菌肽高效分泌株,摇瓶培养确定分泌表达最适p H值。[结果]获得Cath-BF的毕赤酵母高效组成型分泌表达株,表达产物可有效抑制痤疮丙酸杆菌的生长,抗菌肽分泌水平与培养基p H值高度相关。[结论]成功构建金环蛇抗菌肽Cath-BF组成型高效分泌表达系统。

英文摘要：

[ Objective]To establish the efficient secretory expression system of krait antibacterial peptide Cath- BF in Pichia pastoris. [Methods]Cath- BF coding sequence was synthesized by overlaping extension PCR, and cloned into pGAPZa- A. Pichia pastoris SMDl168 was employed as the host and transformed by electroporation,from which ＂high copy＂ transformants were screened with zeocin and selected as the recombinant P. pastoris for secreting Cath - BF. The antibacterial activity of Cath - BF was detected with Propionibacte- rium aches, and the pH optimum was determined by antibacterial activity of supernate from recombinant cells hake - flask cultured in YPD. [ Results] Cath -BF was constitutively expressed and secreted by recombinant P. pastoris, the expression product can effectively inhibited the growth of P. aches. The secretion level of antibacterial peptide has a high correlation with the pH value of culture medium. [ Conclu- sion ] A high efficient constitutively secretory expressing system for antibacterial peptide Cath -BF was established based on P. pastoris.