生长分化因子15基因过表达慢病毒载体的构建

摘要

目的：构建生长分化因子15（growth differentiation factor 15，GDF15）基因过表达慢病毒载体。方法根据GDF15 mRNA 的基因序列，合成GDF15基因，构建至过表达载体并转化至感受态细胞，再进行测序验证。重组慢病毒载体经过测序鉴定后，转染293T 细胞，包装生产慢病毒。用慢病毒转染293T 细胞，再用Western blot 检测GDF15的表达情况。结果测序结果证实GDF15基因正确插入载体中。慢病毒转染293T 细胞后，GDF15基因在蛋白质水平上表达显著增加。结论 GDF15基因过表达慢病毒载体构建成功，并有效增强GDF15基因在293T 细胞中的表达。%Objective To construct lentiviral vectors of GDF15 gene overexpression. Methods Based on the mRNA sequence of GDF15 gene,the GDF15 gene was made. The overexpression vectors were constructed and transfected into competent cells which were detected by sequencing. The 293T cells were transfected by the recombinant lentiviral vectors to achieve the lentivirus packaging pro-duction. The lentiviruses were transfected into 293T cells. The expression of GDF15 gene in 293T cell was detected by Western blot in order to determine the validity of GDF15 gene overexpression. Results It was confirmed by sequencing that the GDF15 gene was cor-rectly inserted into the vectors,and that the GDF15 gene overexpressed lentiviral vectors were successfully constructed. After the lenti-viruses transfected 293T cells,the expression of GDF15 gene increased significantly at protein level. Conclusions The lentiviral vec-tors of GDF15 gene overexpression were constructed successfully,and they efficiently up-regulated the expression of GDF15 in 293T cells.