猪PR-39抗菌肽基因在毕赤酵母中的表达  

Expression of pig PR-39 antibacterial peptide gene in Pichia pastoris

作　　者：江学斌[1] 陈嘉蔚[2] 杨军[2] 柯德森[1] 肖安吉 卢志鹏[2] 楚品品 黄朝远 蔡海明[2] 马苗鹏[2] 张玲华[2] 

机构地区：[1]广州大学生物工程研究所,广东广州510006 [2]华南农业大学生命科学学院广东省农业生物蛋白质功能与调控重点实验室,广东广州510642

出　　处：《生物技术》2016年第1期29-33,47,共6页Biotechnology

基　　金：广州市属高校科技计划项目(No.1201420755)资助

摘　　要：[目的]在毕赤酵母中表达抗菌肽PR-39基因,获得有抗菌活性的PR-39。[方法]根据酵母和猪密码子偏好性,对其密码子进行优化改造。将经SOE-PCR获得的PR-39基因与毕赤酵母表达载体pPIC9K连接,构建重组载体pPIC9K-PR-39。经SacⅠ线性化电击转化毕赤酵母GS115,取阳性克隆进行髙拷贝转化子筛选和诱导表达。[结果]pPIC9K-PR-39重组质粒构建成功,pPIC9K-PR-39菌株发酵产物检测结果对DH5α大肠杆菌和金黄色葡萄球菌都有抑菌效果。[结论]获得了PR-39基因的重组酵母,并用毕赤酵母系统成功地分泌表达了具有明显抗菌活性的抗菌肽PR-39。[ Objective] Expression of pig antimicrobial peptide PR -39 gene in Pichia Pastoris, and obtaining PR -39 with antimicrobial activity. [ Methods ] According to the partiality codon of yeast and pig, the codons were optimized. The PR - 39 gene was amplified by SOE - PCR and ligated to pPIC9K to construct the recombinant expression vector pPIC9K - PR - 39. The recombinant vector was linearized by Sac I ,and transformed into GSll5 by electroporation. The positive clones were screened for high copy transformants and induced by methanol. [ Results] The recombinant carrier pPIC9K -PR -39 has been constructed, and the detection of pPICgK - PR - 39 strain fermentation products shows that the PR - 39 peptide has the antibacterial activity against E. coli and S. aureus. [ Conclusion ] The PR - 39 recombinant yeast has been obtained and the antibacterial peptide PR - 39 with obvious activity has been successfully expressed in Pichia pastoris system.

关 键 词：抗菌肽 PR-39 毕赤酵母 表达 抑菌活性 

分 类 号：Q782[生物学—分子生物学]