猪带绦虫六钩蚴HSP的克隆和表达  

Cloning and Expression of HSP of Taenia Solium Oncosphere

作　　者：王哲[1] 赵权[2] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,大庆163319 [2]吉林农业大学动物科学技术学院

出　　处：《黑龙江八一农垦大学学报》2013年第3期34-37,66,共5页journal of heilongjiang bayi agricultural university

基　　金：国家自然科学基金(NO.30972176)

摘　　要：提取猪带绦虫激活和未激活六钩蚴总RNA,RT-PCR扩增HSP目的基因,将目的基因与pGH克隆载体连接,经酶切鉴定后,将阳性重组质粒进行测序,结果扩增出激活的六钩蚴的目的片段。将目的基因亚克隆到原核表达载体pET-28a-HSP中,并将获得的pET28a-HSP阳性重组子转化至宿主菌E.coliBL21,IPTG进行诱导表达,并对重组抗原pET-28a-HSP进行SDS-PAGE和Western-blot检测。结果表明重组经SDS-PAGE分析可见一条约35 kDa大小的融合蛋白条带的抗原,Western-blot结果显示其能被囊虫病人阳性血清识别。这将为进一步阐明六钩蚴入侵中间宿主的机理、设计新型抗猪囊虫病和绦虫病疫苗打下基础。The HSP gene was separately amplified from total RNA of activated Taenia solium oncosphere by RT-PCR.The PCR products were cloned into pGH vector, recombinant positive clones was sequenced after restriction enzyme digestion.The HSP gene was subcloned into pET28a expression vector,the recombinant pET28a-HSP infected into E.coliBL21.IPTG was added to induce fusion expression and the expression products was identified by SDS-PAGE and Western-blot.One fusion protein band about 35 kDa was identified by SDS-PAGE after inducible expression after inducible expression,The resuh would lay foundations for the mechanism of invasion of between oncosphere and host, and the design of new vaccine anti-porcine cysticercosis and taeniasis.

关 键 词：猪带绦虫六钩蚴 HSP基因 克隆 表达 

分 类 号：S855.9[农业科学—临床兽医学]