短小乳杆菌重组S-层蛋白质的纯化及其体外黏附性的初步鉴定  

Purification of Recombinant S-layer Protein fromLactobacillus brevis and Preliminary Identification of Adhesive Property in Vitro

作　　者：王彩凤[1] 敖敦格日勒[1] 小琴[1] 特尼格尔[1] 赵慧[1] 格日勒图[1] 

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018

出　　处：《动物医学进展》2013年第7期57-61,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(30960279);内蒙古自然科学基金项目(2011BS0408)

摘　　要：以本实验室已构建的pMD19T-slp为模板,应用PCR方法亚克隆slp基因,将其定点插入到原核表达载体pGEX-4T-3中,成功构建原核表达载体pGEX-slp,经IPTG诱导获得重组融合蛋白GST-SLP并用LiCl的方法进行纯化。应用SDS-PAGE和Western blot方法对表达产物进行鉴定,应用ELISA方法鉴定重组融合蛋白的体外黏附特性。在大肠埃希菌裂解样品中,SDS-PAGE和Western blot结果均证明分子质量大小约为71ku的重组融合蛋白GST-SLP的存在;ELISA结果提示,该重组融合蛋白在体外能够有效地黏附在乳酸乳球菌NZ9000表面。本试验为S-层蛋白质生物学特性的进一步研究奠定了基础。Slp gene was subcloned by PCR from the plasmid of pMD19T-slp constructed in our laboratory,and inserted into the prokaryotic expression vector of pGEX-4T-3,so the prokaryotic expression vector of pGEX-slp was successfully constructed.Recombinant fusion protein of GST-SLP was gained by IPTG induction and purified by LiCl method.Then,expression products were identified by SDS-PAGE and Western blot.And the adhesive property of recombinant fusion protein was identified by ELISA method invitro.In cracked samples of E.coli BL21,both the SDS-PAGE and Western blot proved that the presence of the fusion protein GST-SLP which had a molecular weight of approximately 71ku.The ELISA results suggested that the recombinant fusion protein could effectively adhere to the surface of Lactococcus lactis NZ9000 in vitro.These experiments lay a foundation for further study on the biological characteristics of the S-layer protein.

关 键 词：短小乳杆菌 重组S-层蛋白质 纯化 体外 黏附特性 

分 类 号：Q789[生物学—分子生物学] Q51