肺癌细胞A549抗原相关旋毛虫Tsp06172基因的克隆及原核表达  

Cloning and prokaryotic expression of the Tsp06172 gene of Trichinella spiralis in A549 lung cancer cells

作　　者：高江明[1] 徐晓芳[1] 吕萌[1] 左绍志[1] 宫鹏涛[1] 杨举[1] 李赫[1] 李建华[1] 张国才[1] 张西臣[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《中国病原生物学杂志》2013年第3期245-247,共3页Journal of Pathogen Biology

基　　金：国家973计划项目(No.2006CB910505);吉林省科技发展计划项目(No.2003055025;No.20106044)

摘　　要：目的克隆旋毛虫(Trichinella spiralis)与肺癌细胞A549相关抗原Tsp06172基因,并进行原核表达。方法采用RT-PCR方法扩增Tsp06172基因,连接原核表达载体pET-28a,转化入感受态细胞BL21,IPTG诱导表达,经SDS-PAGE和Western blot鉴定表达产物。结果重组表达质粒经双酶切及测序鉴定正确。表达分子质量单位约为16ku的融合蛋白。Western blot检测融合蛋白能被抗A549细胞的多克隆抗体识别。结论构建的原核表达载体pET-28a-Tsp06172表达具有A549细胞反应原性的蛋白,为旋毛虫Tsp06172重组蛋白功能的研究了奠定基础。Objective To clone and express the Trichinella spiralis Tsp06172 gene in BL21. Methods TheTsp06172 gene was amplified with RT PCR and then subcloned into the prokaryotic expression vector pET-28a. BL21 containing the recombinant plasmid pET-28a-Tsp06172 was induced with IPTG. The fusion protein was detected and identified with SDS PAGE and Western blotting. Results The recombinant expression plasmid was successfully constructed. After induction in an E. coli system, SDS PAGE results showed that a fusion protein of about 16 ku was successfully expressed. Western blotting indicated that the fusion protein was readily recognized by polyclonal antibodies from A549 cells. Conclusion The recombinant expression plasmid pET-28a-Tsp06172 expressed the corresponding protein in BL21. This finding lays the foundation for research into the function of the Tsp06172 protein.

关 键 词：旋毛虫 Tsp06172基因 原核表达 肺癌细胞A549 

分 类 号：R383.15[医药卫生—医学寄生虫学]