与乳腺癌细胞MCF-7相关的旋毛虫TS498基因的克隆与表达  

Cloning and expression of TS498 gene of breast cancer MCF-7 cells-associated Trichinella spiralis

作　　者：任保彦[1] 左绍志[1] 高江明[1] 付成福[1] 张旭[1] 宫鹏涛[1] 李建华[1] 张西臣[1] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062

出　　处：《中国生物制品学杂志》2012年第3期300-303,共4页Chinese Journal of Biologicals

基　　金：吉林省科技发展计划项目(2003055025;20106044);国家973课题(2006CB910505;2010CB530004)

摘　　要：目的克隆与乳腺癌MCF-7细胞相关的旋毛虫TS498基因,并进行原核表达。方法从含TS498基因的噬菌体文库中PCR扩增TS498基因,亚克隆入原核表达载体pET-28a(+)中,构建重组表达质粒pET-28a-TS498,转化大肠杆菌BL21(DE3),IPTG诱导表达,SDS-PAGE和Western blot分析表达产物。结果扩增的旋毛虫TS498基因与GenBank中登录的基因序列同源性为100%,重组表达质粒pET-28a-TS498经双酶切和测序证明构建正确;表达的重组蛋白相对分子质量约23 000,表达量约占菌体总蛋白的20%,能够被兔抗旋毛虫肌幼虫阳性血清识别。结论成功克隆并在大肠杆菌中表达了与MCF-7细胞相关的旋毛虫TS498基因,为进一步研究其特性和功能奠定了基础。Objective To clone the TS498 gene of breast cancer MCF-7 cells-associated Trichinella spiralis and express in prokaryotic cells.Methods TS498 gene was amplified from phage library by PCR and subcloned into prokaryotic expression vector pET-28a（＋）.The constructed recombinant plasmid pET-28a-TS498 was transformed to E.coli BL21（DE3） and induced with IPTG.The expressed product was identified by SDS-PAGE and Western blot.Results The homology of amplified TS498 gene sequence was 100% to that reported in GenBank.Restriction analysis and PCR proved that recombinant plasmid pET-28a-TS498 was constructed correctly.The expressed recombinant protein,with a relative molecular mass of about 23 000,contained about 20% of total somatic protein and was recognized by rabbit antiserum against muscle larvae of T.spiralis.Conclusion The TS498 gene of MCF-7 cells-associated T.spiralis was successfully cloned and expressed in E.coli,which laid a foundation of further study on the character and function of the gene.

关 键 词：旋毛虫 TS498基因 克隆 原核细胞 基因表达 乳腺癌 

分 类 号：R383.15[医药卫生—医学寄生虫学] R737.9[医药卫生—基础医学]