重组家蝇抗菌肽Des-HF在酵母中的表达及对金黄色葡萄球菌的抗菌活性  

Secrective expression of recombinant antibacterial peptide Des-HF in Pichia pastoris and its antibacterial activity against Staphylococcus aureus

作　　者：仇妍虹[1,2] 李鹏[1] 李斐[3] 郑其升[1] 曹瑞兵[1] 周斌[1] 陈德胜[1] 陈溥言[1] 

机构地区：[1]南京农业大学农业部动物疫病诊断与免疫重点开放实验室,江苏南京210095 [2]北京市兽医卫生监督所,北京100044 [3]青岛农业大学生命科学学院,山东青岛266109

出　　处：《南京农业大学学报》2009年第1期105-109,共5页Journal of Nanjing Agricultural University

摘　　要：参考家蝇防御素抗菌肽基因序列(AY260152)设计4条引物,用降落PCR方法获得家蝇防御素抗菌肽成熟肽基因。将基因克隆入酵母表达载体pPICZα-A,构建分泌型重组酵母表达载体pPIC-Des-HF,转化Pichia pastoris受体菌SMD1168,在醇氧化酶(AOX)启动子调控下,相对分子质量约5 000的重组抗菌肽Des-HF获得表达。抗菌试验结果表明,该表达产物对金黄色葡萄球菌有较好的抑菌活性。以小鼠为试验模型研究重组抗菌肽对金黄色葡萄球菌的体内抗菌活性,试验结果表明,240μg重组Des-HF可分别保护小鼠免受10倍最小致死浓度的金黄色葡萄球菌(ATCC26003)和CowanⅠ的攻击。With four pairs of specific primers designed according to the relevant sequence published on GenBank, the housefly antibacterial peptide defense gene was amplified with touch-down PCR method. After sequencing, the target gene was directly cloned into multiple cloning site of the pastoris expression plasmid pPICZα-A to get the recombinant plasmid pPIC-Des-HF. The 5 000 relative molecular weight recombinant antibacterial peptide Des-HF was seerectively expressed in the Pichia pastoris host cell SMD1168, when induced with methanol. The recombinant Des-HF showed perfect bacteriostasis activity against Gram-positive bacteria. Mice as animal model were used to test bacteriostasis activity and the result indicated that 240 μg recombinant Des-HF could completely protect the mice from ten folds minimum lethal dose concentration of Staphylococcus aureus （ ATCC26003 ） and Cowan Ⅰ.

关 键 词：家蝇防御素抗菌肽 Des—HF 酵母 分泌表达 金黄色葡萄球菌 抗菌活性 

分 类 号：S852.44[农业科学—基础兽医学]