小鼠睾丸基因PDZD9的克隆及亚细胞定位分析  

Cloning and Subcellular Location of a Novel Mouse Testis Gene PDZD9

作　　者：周军媚[1] 姚峰[2] 刘鑫[2] 王宗保[2] 吴端生[2] 陈春芳[2] 贺震[2] 余坚[2] 

机构地区：[1]南华大学药学与生命科学学院,湖南衡阳421001 [2]南华大学实验动物学部

出　　处：《中南医学科学杂志》2012年第2期127-130,共4页Medical Science Journal of Central South China

基　　金：国家自然科学基金资助项目(81000035);湖南省科技厅项目(湘财教指[2008]115)

摘　　要：目的克隆小鼠睾丸基因PDZD9并构建pEGFP-C1-PDZD9重组载体进行亚细胞定位分析。方法提取小鼠睾丸RNA,利用逆转录—聚合酶链反应(RT-PCR)技术从小鼠睾丸cDNA中分离PDZD9基因。将PDZD9基因定向克隆至带有绿色荧光报告基因的真核表达载体pEGFP-C1。利用阳离子聚合物方法转染COS7细胞,荧光显微镜下观察其亚细胞定位。结果小鼠睾丸基因PDZD9克隆成功,成功构建真核表达载体pEGFP-C1-PDZD9,亚细胞分析表明该蛋白定位于细胞核和细胞质中,主要分布在细胞核中。结论成功克隆了小鼠睾丸基因PDZD9,并初步分析了其在细胞中的定位情况,为下一步对PDZD9的功能研究奠定了基础。Objective To clone a novel testis gene PDZD9,construct the eukaryotic expression vector pEGFP-C1-PDZD9 and analysis subcellular location of PDZD9 in COS-7 cell.Methods Total RNA was extracted from the testes of normal mature mouse,PDZD9 gene was amplified by RT-PCR and inserted into the eukaryotic expression vector pEGFP-C1.The recombinant plasmid was transfected into COS-7 cells by cationic polymer.Observation under the fluorescent microscopy was used to analysis the subcellular location of PDZD9.Results The mouse gene PDZD9 was cloned and the eukaryotic expression vector pEGFP-C1-PDZD9 was constructed successfully.Observation under the fluorescent microscopy showed that PDZD9 was expressed in cytoplasm and nucleus,mainly in cell nucleus.Conclusions The gene cloning of PDZD9 was successful and initially proved the subcellular location of PDZD9,which would be benefited for further investigation on the function of PDZD9 gene.

关 键 词：PDZD9 克隆 转染 亚细胞定位 

分 类 号：R541.4[医药卫生—心血管疾病]