人NR2B基因真核表达载体的构建及其在CHO细胞中的表达

目的 克隆人NR2B基因,构建其真核表达载体,获得暂态表达NR2B的CHO细胞.方法 RT-PCR方法克隆人NR2B基因,并插入真核表达载体pcDNA3.1中,将该重组载体转染至CHO细胞.通过RT-PCR、Western blot及间接免疫荧光鉴定细胞中NR2B的表达,通过流式细胞仪检测细胞的凋亡.结果 成功获得人NR2B基因,转染的CHO细胞可检测到NR2B的表达,表达NR2B的CHO细胞并不会凋亡.结论 成功克隆和构建了人NR2B基因的真核表达载体,并在CHO细胞中得到了表达.

Construction of human NR2B eukaryotic expression vector and its expression in CHO cells

Objective To clone human NR2B gene, construct its eukaryotic expression vector, and temporarily express it in CHO cells. Methods Human NR2B gene was amplified by RT-PCR and then inserted into eukaryotic vector pcDNA3.1. The recombinant plasmid was transfected into CHO cells. The expression of the target molecule was identified by RT-PCR, Western blotting, indirect immanofluorescent staining and the apoptosis was detected by flow cytometry. Results The NR2B gene was obtained; after transfection, NR2B was successfully expressed in CHO cells, and the expression of NR2B did not induce the apoptosis of CHO cells. Conclusion Human NR2B gene has been successfully cloned and expressed in CHO cells via constructing its eukaryotic expression vector.