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目的克隆人CD160基因,并构建含有该目的基因的pIRES2-EGFP重组质粒载体,获得稳定表达CD160分子的基因转染细胞。方法从人外周血cDNA文库中扩增CD160的基因,另以VSIG4基因的cDNA为模板扩增跨膜区基因片段,将它们一并装入pIRES2-EGFP质粒载体中,脂质体法共转染L929细胞,用G418筛选出能稳定表达CD160分子的L929细胞株。结果成功克隆出了CD160的基因,构建了pIRES2-EGFP/CD160TMV质粒载体,并建立了稳定表达人CD160分子的L929转基因细胞株,该转基因细胞能结合L929/HVEM转基因细胞,证明其功能性。结论构建了含人CD160基因的重组pIRES2-EGFP质粒载体和建立稳定表达人CD160分子的细胞株,为CD160分子的后续研究奠定基础。
Objective To clone human CD160 gene and construct pIRES2-EGFP recombinant plasmid vector containing the target gene to obtain the gene transfection cells stably expressing CD160. Methods CD160 gene was amplified from human peripheral blood cDNA library. The cDNA of VSIG4 gene was used as a template to amplify transmembrane region gene fragments, which were then inserted into pIRES2-EGFP plasmid vector and co-transfected by liposome L929 cells. The L929 cell line stably expressing CD160 was screened by G418. Results The CD160 gene was successfully cloned and the pIRES2-EGFP / CD160TMV plasmid vector was constructed. The L929 transgenic cell line stably expressing the human CD160 gene was established. The transgenic cell line can bind to the L929 / HVEM transgenic cell to prove its functionality. Conclusion The recombinant plasmid pIRES2-EGFP containing human CD160 gene and the cell line stably expressing human CD160 were constructed, which laid the foundation for the subsequent study of CD160.