AQP1真核表达载体的构建及其在FRT细胞的表达  

Construction of Eukaryotic Expression Vector for AQP1 and Its Expression in FRT Cells

作　　者：张引红[1] 刘田福[1] 冯学超[2] 苏惟恒[2] 麻彤辉[2] 

机构地区：[1]山西医科大学实验动物中心,山西太原030001 [2]东北师范大学膜通道实验室,吉林长春130024

出　　处：《激光生物学报》2010年第4期475-478,474,共5页Acta Laser Biology Sinica

基　　金：国家自然科学杰出青年基金项目(30325011)

摘　　要：目的:构建小鼠AQP1基因真核表达质粒并观察其在FRT细胞中的表达。方法:采用RT-PCR方法从小鼠肾脏组织的总cDNA中扩增出小鼠的AQP1基因,采用基因重组技术将AQP1的cDNA片段插入真核表达载体pCAGGS,构建小鼠AQP1的真核表达质粒,脂质体转染FRT细胞进行表达。结果:酶切和测序结果证实AQP1真核表达质粒构建成功,经脂质体转染FRT细胞后,免疫荧光检测证明AQP1蛋白在真核细胞中成功表达。结论:成功构建真核表达质粒pCAGGS-AQP1-myc,并在FRT细胞中得以表达。为进一步研究小鼠AQP1过表达时的功能及机制奠定了实验基础。Objective： To construct a recombinant mouse AQP1 eukaryotic vector and observe its expression in FRT cell in vitro. Methods：AQP1 gene amplified by RT-PCR from mouse kidney tissue was inserted into eukaryotie expression vector pCAGGS,then the recombinant pCAGGS-AQP1 plasmid was transfected into FRT cells using Lipofectamine. Re- suits： AQP1 eukaryotie vector was successfully constructed and confirmed by endonuclease digestion and DNA sequencing. The expression of AQP1 protein was detected by immunofluorescence,which suggested that it could be expressed in eukaryotic cells. Conclusion： These results provided a basis for further investigation of AQP1 function.

关 键 词：水通道蛋白1 载体构建 表达 

分 类 号：Q786[生物学—分子生物学]