重组穿梭质粒pAdTrack-CMV-PGC1α的构建及鉴定  

Construction and identification of recombinant plasmid pAdTrack-CMV-PGC1 encoding the mouse PGC1α gene

作　　者：马仕坤[1] 程庆丰[1] 青华[1] 冯正平[1] 李卫平[1] 周波[1] 张素华[1] 李启富[1] 

机构地区：[1]重庆医科大学附属第一医院内分泌科,400016

出　　处：《重庆医学》2006年第14期1249-1250,1268,共3页Chongqing medicine

基　　金：国家自然科学基金资助项目(30570876)

摘　　要：目的构建编码小鼠PGC1α基因的质粒,为进一步研究定向诱导PGC1α在白色脂肪组织特异性表达做准备。方法提取小鼠肾脏总RNA,利用RT-PCR扩增目的基因,引物设计时在两端分别引入EcoRV和SalⅠ的酶切位点,双酶切后与经过同样处理的穿梭质粒载体pAdTrack-CMV相连,经酶切和PCR鉴定。结果通过RT-PCR获取了目的基因并成功克隆入载体,目的基因序列与GENEBANK报道序列一致。结论构建出编码小鼠PGC1α基因的重组质粒pAdTrack-CMV-PGC1α。Objective To construct a plasmid encoding mouse PGC1α gene, for the further research on the aP2 promoter/enhancer-driven mouse PGC1α tissue-specific expression in white adipose tissue. Methods The complementary DNA of the mouse PGC1α gene was amplified by RT-PCR from the C57 mouse kidney and cloned into the pAdTrack-CMV vector to obtain the plasmid pAdTrack-CMV-PGC1α and then identified by enzymatic cutting and PCR products sequencing. Results The mouse PGC1α gene was amplified through RT-PCR and cloned into the transfer vector. The favorite gene sequence was completely consistent with that reported in GENEBANK. Conclusion The recombinant plasmid pAdTrack-CMV-PGC1α encoding PGC1α gene was successfully constructed.

关 键 词：PGC1α 脂肪组织 穿梭质粒 

分 类 号：R589.2[医药卫生—内分泌]