生长分化因子15基因过表达慢病毒载体的构建  

Construction of lentiviral vectors of GDF15 gene overexpression

作　　者：王宁涛[1] 张晓晨[2] 张文杰[3] 华洪飞 张志愿[2] 王绍义[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院口腔外科,上海市口腔医学重点实验室,上海200011 [2]上海交通大学医学院附属第九人民医院口腔颌面头颈肿瘤科,上海市口腔医学重点实验室,上海200011 [3]上海交通大学医学院附属第九人民医院修复科,上海市口腔医学重点实验室,上海200011

出　　处：《口腔医学》2015年第9期710-713,共4页Stomatology

基　　金：国家自然科学基金(81271114)

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GDF15mRNA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.

关 键 词：生长分化因子15 过表达 慢病毒载体 

分 类 号：R393[医药卫生—基础医学]