大鼠Tmub1基因过表达慢病毒载体的构建  

Construction of lentivirus vectors of rat Tmub1 gene overexpression

作　　者：赵晓彪[1] 李光耀[1] 刘孟刚[1] 范霞[2] 陈平[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所肝胆外科,重庆400042 [2]第三军医大学大坪医院野战外科研究所一室,重庆400042

出　　处：《重庆医学》2016年第13期1744-1746,共3页Chongqing medicine

基　　金：国家自然科学基金面上项目(81270523)

摘　　要：目的构建Tmubl基因的过表达慢病毒载体（LV—Tmubl），为研究Tmubl蛋白在肝细胞增殖过程中的作用提供实验材料。方法化学合成Tmubl基因序列，用BamHI／AgeI酶切化学合成含有目的基因的质粒及GV287载体，PCR产物连接入线性化表达的载体。PCR鉴定引物，再对PCR鉴定阳性的克隆进行DNA测序和比对分析。使用构建的LV—Tmnbl，转染293T细胞，荧光法检测构建的慢病毒滴度。结果成功构建了LV-Tmubl，并获得相应的病毒，病毒滴度为2×10^8TU／mL。结论LV-Tmubl为进一步研究Tmubl蛋白在肝细胞增殖中的作用提供了实验基础。Objective To construct the lentiviral vector with overexpression Tmubl gene to provide the experimental materials in the research of Tmubl protein function in the process of the hepatocyte proliferation. Methods The Tmubl gene sequences was constructed through chemical synthesis,plasmid containing the purpose gene and GV287 vectors were digested with BamHI/ AgeI enzyme, the PCR products were connected into the linearized expression vector. PCR confirmed the primers,the positive clones identified by PCR DNA were sequenced and performed the comparative analysis. The constructed lentiviral vectors of Tmubl gene overexpression was used to transfect 293T cell lines. Then the lentivirus titer was detected by the fluorescence method. Results The lentiviral vector with Tmubl gene overexpression was successfully constructed and the corresponding virus was obtained,the virus titer was2×10^8 TU/mL. Conclusion LV-Tmubl expressing lentiviral vector provides the experimental foundations for the further study of the role of Tmubl protein in hepatocyte proliferation.

关 键 词：Tmub1 基因 过表达慢病毒载体 大鼠 

分 类 号：R575.3[医药卫生—消化系统]