PDX-1表达质粒的构建及表达  

Construction and expression of the PDX-1-expression plasmid

作　　者：夏华强[1] 刘律君[1] 徐小明[1] 卢光琇[1] 

机构地区：[1]中南大学生殖与干细胞工程研究所,人类干细胞国家工程研究中心,湖南长沙410078

出　　处：《现代生物医学进展》2009年第22期4265-4268,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金(30800650);国家973项目(2007CB947900);国家863项目(2006AA02A102)资助

摘　　要：目的:为进一步研究PDX-1基因的功能打下基础。方法:从人的胰腺cDNA中PCR扩增PDX-1基因的阅读框架,构建质粒pAAV-PDX-1。用pAAV-PDX-1转染293T细胞48小时后,用RT-PCR和Western Blot方法检测293T细胞中mRNA和蛋白水平PDX-1的表达。结果:转染了pAAV-PDX-1的293T细胞中有PDX-1的表达,而在转染质粒pAAV-MCS和未转染的293T细胞中均没有检测到PDX-1的表达。结论:构建的pAAV-PDX-1质粒在mRNA和蛋白水平都能表达PDX-1。Objective：To further study the function of the PDX-1 gene. Methods： We directly cloned a 987-bp fragment including PDX-1 open reading frame （ORF） from the human pancreas cDNA to construct pAAV-PDX-1 plasmid. The plasmid was transfected into the 293T cells and the expression of PDX-1 was analyzed by RT-PCR and Western blot analysis. Results： The PDX-1 expression was detected in the 293T cells transfected with pAAV-PDX-1 while not detected in the 293T cells transfected with control plasmid or untransfected. Conclusion： We successfully constructed plasmid pAAV-PDX-1 that expressed the PDX-1, which provide the base to study the function of the PDX-1.

关 键 词：质粒构建 pAAV-PDX-1 293T细胞 

分 类 号：R587.1[医药卫生—内分泌]