铜绿假单胞菌噬菌体PaP1内溶素基因克隆、表达及活性分析  

Cloning,expression and enzymatic assay of bacteriophage PaP1 endolysin against Pseudomonas aeruginosa

作　　者：孙卫忠[1] 胡晓梅[1] 饶贤才[1] 谭银玲[1] 陈志谨[1] 丛延广[1] 胡福泉[1] 

机构地区：[1]第三军医大学基础医学部微生物学教研室,重庆市微生物工程实验室,重庆400038

出　　处：《第三军医大学学报》2008年第13期1268-1271,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30300295)~~

摘　　要：目的构建铜绿假单胞菌噬菌体内溶素重组原核表达载体,检测重组蛋白的酶学活性及抑菌活性。方法利用重组技术将铜绿假单胞菌裂解性噬菌体PaP1内溶素基因构建到表达载体pQE31中,并在大肠杆菌M15(pREP4)中诱导表达,通过非变性方法纯化重组融合蛋白,利用酶谱电泳、抑菌实验等技术检测酶学活性及抑菌活性。结果酶谱电泳方法结果显示重组融合蛋白对铜绿假单胞菌细胞壁肽聚糖有降解作用,抑菌活性检测结果显示重组融合蛋白对金黄色葡萄球菌有一定的抑菌效果。结论该研究从实验方面证实了噬菌体PaP1内溶素基因的生物学功能。Objective To construct, express and purify recombinant endolysin of Pseudomonas aeruginosa the expression vector pQE31. Then the fusion protein expressed in E. coli M15 （pREP4） and purified under native conditions. The enzymatic activity was detected by zymography assay and its biological activity was detected by antibacterial activity assay. Results Enzymatic assay showed that the recombinant protein could cause the lysis of the purified P. aeruginosa peptidoglycan. Antibacterial activity assay showed that the recombinant protein inhibited the growth of Staphylococcus aureus. Conclusion The obtained phage endolysin exerts antibacterial activity to P. aeruginosa.

关 键 词：噬菌体 内溶素 基因表达 活性鉴定 

分 类 号：Q939.48[生物学—微生物学] R378.991[医药卫生—病原生物学]