两歧双歧杆菌中serpin基因的克隆、表达与功能分析  

Cloning,Expression and Function Analysis of Serpin Gene in Bifidobacterium bifidum

作　　者：叶若松[1] 黎鹏[1] 章昭琳[1] 万翠香[1] 崔佳[1] 魏华[1] 

机构地区：[1]南昌大学食品科学与技术国家重点实验室,南昌330047

出　　处：《中国生物工程杂志》2011年第2期38-42,共5页China Biotechnology

基　　金：国家"863"计划(2008AA10Z337);国家自然科学基金(30900038);江西省主要学科学术带头人培养计划(2009)资助项目

摘　　要：从Bifidobacterium bifidum WBBI02基因组中克隆了serpin基因片段,构建了重组Serpin蛋白的原核表达体系,实现了Serpin的表达与纯化。纯化的Serpin蛋白进行了抑制肠道蛋白酶活性检测,以及对双歧杆菌粘附作用影响的显微观察研究。结果表明:WBB I02中长度为768 bp的serpin基因序列,与GENEBANK中Bifidobacterium longum NCC2705serpin序列同源性为99.9%。原核表达载体pBX2-WBBI02表达的Serpin能有效地抑制糜蛋白酶和胰弹性蛋白酶的活性,最高抑制率分别为90%和97%,显微观察结果证实Serpin能促进双歧杆菌对HT-29细胞的粘附。The Serpin gene has been cloned from the genomic DNA of Bifidobacterium bifidum WBB102. A recombinant Serpin expression system in prokaryotic cells, namely, pBX2- WBB102 was constructed, and the Serpin protein was successfully expressed and purified. An in vitro inhibition test of Serpin against intestinal proteinase and the effect of Serpin on the adherence of Bifidobacterium longum were approached. The results showed that the serpin gene of Bifidobacterium bifidum WBBI02 is 768 bp, whose similarity is 99.9% with that of Bifidobacterium longum NCC2705. The purified Serpin was efficient to inhibit the activities of eukaryotic α- chymotrypsin and pancreatic elastase by maximum 90% and 97%, respectively, microscopical observation proved that Serpin enhanced the adherence of Bifidobacteria to HT-29 cells.

关 键 词：双歧杆菌 SERPIN 克隆表达 粘附 

分 类 号：Q789[生物学—分子生物学]