论文部分内容阅读
目的构建表达人Sprouty 2(SPRY2)基因的重组表达载体pDC315/hSPRY2,转染人胚肾(HEK)293T细胞表达目的蛋白,初步探讨其对HEK293T细胞增殖与存活的影响。方法构建hSPRY2重组腺病毒表达载体,体外转染HEK293T细胞,以Western blot法检测目的蛋白的表达,以CCK-8法检测SPRY2对HEK293T细胞增殖或去血清条件下细胞存活的影响。结果成功构建重组表达载体pDC315/hSPRY2,在转染的HEK293T细胞中检测到目的蛋白hSPRY2的表达,转染pDC315/hSPRY2组HEK293T细胞的增殖率及存活率均明显高于对照组即转染pDC315/EGFP组(P<0.05)。结论成功构建了hSPRY2重组表达载体,过表达hSPRY2可促进HEK293T细胞的增殖与存活。
Objective To construct the recombinant expression vector pDC315 / hSPRY2 expressing human Sprouty 2 gene and to express the target protein in HEK293T cells. The aim of this study was to investigate the effect of SPRY2 on the proliferation and survival of HEK293T cells. Methods The hSPRY2 recombinant adenovirus expression vector was constructed and transfected into HEK293T cells in vitro. The expression of the target protein was detected by Western blot. The effect of SPRY2 on the proliferation of HEK293T cells or the survival of the cells were detected by CCK-8. Results The recombinant plasmid pDC315 / hSPRY2 was successfully constructed and the expression of hSPRY2 protein was detected in HEK293T cells transfected with pDC315 / hSPRY2. The proliferation and survival rates of HEK293T cells transfected with pDC315 / hSPRY2 were significantly higher than those of the control cells transfected with pDC315 / EGFP group (P <0.05). Conclusion The hSPRY2 recombinant plasmid was successfully constructed and hSPRY2 was overexpressed to promote the proliferation and survival of HEK293T cells.