论文部分内容阅读
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 Trichinella spiralis and lung cancer cell A549 antigen Tsp06172 gene and prokaryotic expression. Methods The Tsp06172 gene was amplified by RT-PCR and inserted into prokaryotic expression vector pET-28a. The recombinant plasmid was transformed into BL21 and induced by IPTG. The expressed product was identified by SDS-PAGE and Western blot. Results The recombinant plasmid was identified by double enzyme digestion and sequencing. The expression of the molecular mass unit of about 16ku fusion protein. Western blot detection of fusion protein can be anti-A549 cell polyclonal antibody recognition. Conclusion The prokaryotic expression vector pET-28a-Tsp06172 expressed A549 cell-reactive protein, which laid the foundation for the study of the function of Trichinella spiralis Tsp06172 recombinant protein.