仿生鲶鱼抗菌肽编码序列的设计及克隆

Design and Cloning of an Emulated Sequence of Catfish Parasilurus asotus Antimicrobial Peptide

人工合成的仿生鲶鱼抗菌肽DNA序列,经聚合酶链式反应(PCR)扩增后得到带有不同限制性酶切位点的序列XH1、XH2、XH3.将此基因克隆至载体pGEX-5X-3,成功构建了三串联的重组质粒.测序验证后转化大肠杆菌BL21(DE3),37℃IPTG诱导,获得高效表达,SDS-PAGE扫描分析表明,融合蛋白可达细菌全蛋白总量的30%,融合蛋白以包含体的形式存在于大肠杆菌细胞中.

Sequences XH1、XH2、XH3 coding for antimicrobial peptide with different restriction sites were obtained by PCR amplification,and then cloned into the fusion vector pGEX-5X-3 . The recombinant with three sequence segments was constructed and transformed into E. coli BL21 (DE3) after the expected sequences were checked. Induced by IPTG in 37℃,the cloned protein expressed in a high level and reached about 30% of the total bacterial protein from desitometric scan analysis of SDS-PAGE. The fusion protein existed in E. coli cells was in the form of inclusion body.