类鼻疽伯克霍尔德菌Ⅲ型分泌系统蛋白BsaL的制备和免疫反应性分析  

Expression and immunological evaluation of BsaL,type Ⅲ secretion system protein of Burkholderia pseudomallei

作　　者：方瑶[1] 胡艺[1] 朱攀[1] 任春艳[1] 马腾飞[1] 王昆[1] 毛旭虎[1] 

机构地区：[1]第三军医大学西南医院医学检验系临床微生物及免疫学教研室,重庆400038

出　　处：《第三军医大学学报》2016年第6期585-588,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81471914)~~

摘　　要：目的构建表达重组的类鼻疽伯克霍尔德菌Ⅲ型分泌系统结构蛋白Bsa L的大肠工程菌,制备高纯度、高质量的重组Bsa L蛋白样品,探讨其作为类鼻疽菌检测的候选抗原的可行性。方法基因工程技术构建表达重组Bsa L蛋白的大肠工程菌,通过亲和层析、凝胶过滤层析等纯化方法得到高纯度的Bsa L蛋白。包被Bsa L蛋白于96孔板,分析其用于检测临床类鼻疽菌株的敏感性和特异性。结果成功构建了表达重组Bsa L蛋白的大肠工程菌,诱导、表达及纯化后得到高纯度(>99%)、高浓度(20 mg/m L)的重组Bsa L蛋白。该蛋白在类鼻疽菌检测方面表现出较好的灵敏度(85%)和特异性(89%)。结论成功表达了重组的Bsa L蛋白,获得了高纯度的Bsa L蛋白样品,在类鼻疽菌的诊断方面展示出较好的应用价值。Objective To express recombinant BsaL, type Ⅲ secretion system protein of Burkholderia pseudomaUei, for detecting the bacterium based on enzyme linked immunosorbent assay （ELISA）. Methods PET-28a/BsaL plasmid was constructed and introduced into Escherichia coli BL21. BsaL-His was induced by IPTG and purified by affinity chromatography and gel filtration chromatography. Then, the recombinant protein was applied to 96-well plate for detecting Burkholderia pseudomallei-containing samples. Results BsaL-His with high purity （ 〉99% ） and high concentration （20 mg/mL） was prepared and applied to detecting Burkholderia pseudomaUei at relative high specificity （89%） and sensitivity （85%）. Conclusion BsaL protein is successfully expressed and purified, and it can be applied to detecting Burkholderia pseudomallei.

关 键 词：类鼻疽伯克霍尔德菌 Ⅲ型分泌系统(TTSS) BsaL ELISA 

分 类 号：R341[医药卫生—基础医学] R378.99