细粒棘球蚴重组HSP70基因的表达、纯化及免疫学鉴定  

Expression,Purification and Immunologic Identification of the Recombinant HSP70 Gene From Echinococcus Granulosus

作　　者：于晶晶[1] 于辛酉[1] 王娅娜[1] 师志云[1] 马锐[1] 卜阳[1] 罗永云[1] 赵巍[1] 

机构地区：[1]宁夏医学院基础学院医学遗传学与细胞生物学教研室,银川750004

出　　处：《宁夏医学院学报》2008年第2期140-142,共3页Journal of Ningxia Medical College

基　　金：国家自然科学基金项目(30260105);宁夏自然科学基金项目(NZ0540)

摘　　要：目的构建细粒棘球蚴(Echinococcus granulosus,Eg)热休克蛋白70(Heat shock protein,HSP70)基因的重组质粒并原核表达、纯化及对其免疫特性进行鉴定。方法从重组质粒HSP70/pGEM-T中酶切HSP70目的片段,亚克隆入表达载体pET-28a,转化入大肠杆菌E.coli BL21(DE3)plysS进行融合表达,经His-band树脂纯化试剂盒小量纯化,SDS-PAGE和Western blot方法鉴定表达产物。结果成功构建Eg.HSP70/pET-28a/E.coli BL21基因工程菌株,诱导表达重组蛋白和纯化分离得到14kDaEg.HSP70均能被细粒棘球蚴天然抗原免疫的兔多克隆抗血清识别。结论初步证实该重组蛋白具有较好的抗原性。Objective To const tact HSP70 gene gene and express prokaryotically it, to purify and identify the immunogeuicity of recombinant HSP70 protein. Methods Eg. HSP70 gene was attained from Eg. HSP70/ pGEM-T recombinant plasmid and subcloned into high level expressed vector pET-28a. E.coli BL21 （DE3）plys was sonst mcted. Western blot and ELISA were used to identify the immunogenicity of the recombinant protein. Results E.coli BL21（DE3）plys srecombinant plasmid was const meted. Western blot analysis showed that recombinant protein and protoscolex , cystic fluid and cystic wall of 14kDa Eg. HSP70 could be recognized by this antibody. Conclusion pET-28a/HSP70 recombinant plasmid is constructed and expressed efficiently. The antigenicity and immunogenicity of Eg. HSP70 protein is identified.

关 键 词：细粒棘球蚴 热休克蛋白HSP70 表达 纯化 免疫鉴定 

分 类 号：R383.33[医药卫生—医学寄生虫学]