mTLR2基因在CHO细胞中的表达

Expression of Mouse Toll-Like Receptor 2 in CHO Cell

目的在CHO细胞中表达小鼠TLR2(mTLR2)。方法将目的基因克隆到真核表达载体pcDNA3.1中,并将其转染到CHO细胞,用G418筛选阳性克隆。采用PCR方法,检测外源基因的整合。采用Westernblot、免疫组织化学对转染细胞的mTLR2表达进行鉴定。结果成功构建了pcDNA-mTLR2重组质粒。将其转染到CHO细胞,经G418筛选获得阳性细胞克隆。阳性细胞克隆经PCR检测表明TLR2基因得到稳定整合。Westernblot分析显示在分子量约95000处可见清晰的阳性反应条带。免疫组织化学检测显示转染细胞表达mTLR2。结论获得了表达mTLR2的细胞株,为进一步进行相关研究奠定了基础。

Objective To express mouse toll like receptor 2 （mTLR2） in CHO cell. Methods The mTLR2 gene was cloned into pcDNA3.1 mammalian expression vector, and the recombinant plasmid was transfected into CHO cell by electroporation,then the positive cell clones were selected with G418. The insertion of exogenous gene was analysed by PCR, and expression of mTLR2 in transfected cell was identified by Western blot and immunohistochemical method.Results The mammalian expression plasmid was constructed, and the positive cell clones was obtained. mTLR2 gene was detected in the positive cell clones. One of the positive clones was found to express a protein with a molecular weight of 95 kDa using Western blot analysis. The results suggest that mTLR2 gene was expressed by the positive clones. Conclusion A cell clone expressing mTLR2 was successfully established. It can be used for the further study of TLR.