猪源抗菌肽PBD-1在毕赤酵母中的表达及鉴定  

Expression and Identification of Porcine Antimicrobial Peptide PBD-1 in Pichiapastoris

作　　者：江学斌[1] 陈嘉蔚[2] 杨军[2] 刘顺枝[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,广州510642

出　　处：《中国畜牧兽医》2016年第3期644-649,共6页China Animal Husbandry & Veterinary Medicine

基　　金：广州市属高校科技计划项目(1201420755)

摘　　要：本试验旨在实现猪β防御素1(poricine-β-defensin 1,PBD-1)在毕赤酵母中的表达,获得有抗菌活性的抗菌肽PBD-1。根据PBD-1的氨基酸序列和酵母密码子偏好性,设计优化其核苷酸序列,利用SOE-PCR技术获得PBD-1基因序列,克隆到毕赤酵母表达载体pPIC9K中,构建重组质粒pPIC9K-PBD-1,经SacⅠ线性化后转入毕赤酵母SMD1168中。PCR筛选得到阳性酵母表达菌株,经甲醇诱导后得到分子质量约4.5ku的抗菌肽PBD-1。抗菌特性研究结果表明,表达产物抗菌肽PBD-1对大肠杆菌、金黄色葡萄球菌及枯草芽孢杆菌均有较好的抑制效果。In order to produce antimicrobial peptide PBD-1with bioactivity in Pichia pastoris,according to published amino acid sequence of PBD-1and the partiality codon of yeast,the PBD-1gene was amplified by SOE-PCR and cloned into pPIC9 Kto construct a recombinant expression vector pPIC9K-PBD-1.The recombinant vector was linearized by SacⅠ,and then transformed into SMD1168 by electroporation.Positive yeast expression strain was obtained by PCR.The antimicrobial peptide PBD-1（approximately 4.5ku）was expressed by methanol induction.Antibacterial activity assay showed that the antimicrobial peptide PBD-1had better antibacterial activity against E.coli,S.aureus and B.subtilis.

关 键 词：PBD-1基因 表达 毕赤酵母 PCR 

分 类 号：Q786[生物学—分子生物学]