人p21活化激酶6基因真核表达质粒的构建及其亚细胞定位  

Construction of human PAK6 eukaryotic plasmid and identification of its subcellular localization

作　　者：刘彤[1] 李丹妮[2] 李洋[1] 李丰[1] 

机构地区：[1]中国医科大学基础医学院细胞生物学教研室/教育部医学细胞生物学重点实验室,辽宁沈阳110001 [2]中国医科大学

出　　处：《东南大学学报（医学版）》2013年第3期297-300,共4页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金资助项目(31171360)

摘　　要：目的:构建人p21活化激酶6(PAK6)真核细胞表达质粒,并研究其在人胚肾真核细胞HEK293中的亚细胞定位。方法:PCR扩增人PAK6基因,将其克隆入真核表达载体pcDNA3.1HisC中,构建含人PAK6基因的真核表达质粒pcDNA3.1HisC-PAK6。利用酶切鉴定,DNA测序,免疫印迹及免疫荧光方法鉴定PAK6真核表达质粒及PAK6蛋白的亚细胞定位。结果:经酶切鉴定、DNA测序、免疫印迹方法确认PAK6基因成功导入真核表达载体pcDNA3.1HisC。瞬时转染HEK293细胞,PAK6蛋白表达在细胞浆中。结论:PAK6真核表达质粒构建成功,PAK6定位于细胞浆中。Objective： To construct the eukaryotic pcDNA3.1HisC-PAK6 and identify the subcellular localization of PAK6 in HEK293 cells. Methods： PAK6 fragment was subcloned into pcDNA3.1HisC after PCR.The plasmid was verified by enzyme digestion,DNA sequencing and its expression was subsequently checked by western blot.Immunofluorescence was used to identify its subcellular localization. Results： PAK6 was subcloned into pcDNA3.1HisC sucessfully. Conclusion： Eukaryotic pcDNA3.1HisC-PAK6 was constructed successfully and PAK6 locates in cytoplasm.

关 键 词：PAK6 真核细胞 PCDNA3 1HisC 亚细胞定位 

分 类 号：R736[医药卫生—肿瘤]