代表PDCD5蛋白不同功能区域的质粒构建及其在胃癌细胞中的表达定位  

Construction and expression of plasmids representing different domains of PDCD5 protein in gastric cancer cells

作　　者：王巍[1] 张宁[1] 赵成海[1] 

机构地区：[1]中国医科大学基础医学院病理生理教研室,辽宁沈阳110001

出　　处：《中国现代医学杂志》2012年第17期22-24,共3页China Journal of Modern Medicine

基　　金：辽宁省自然科学基金(No:20092125);辽宁省教育厅高等学校科研项目(No:2009A751)

摘　　要：目的分析PDCD5蛋白的不同功能区域,将这些区域所对应的序列与质粒pEGFP-C1相连接,在胃癌细胞中表达后,检测其细胞定位。方法设计带有HindⅢ和EcoRⅠ酶切位点的PDCD5引物共4对,包括完整的PDCD5蛋白序列及3个不同功能区域的序列。以上质粒构建完成后,转染胃癌细胞SGC-7901。荧光显微镜检测融合蛋白的表达定位。结果经酶切鉴定,可见相应长度的4种序列,后经测序确定序列无误,荧光显微镜检测融合蛋白表达。结论代表PDCD5蛋白不同功能区域的质粒成功构建,并在SGC-7901细胞中表达。经过观察绿色荧光蛋白,确定4种蛋白均定位于细胞浆内。【Objectives】 To analyze the different domains of PDCD5 protein,and connect their sequence to the plasmid of pEGFP-C1.Then to detect their cellular localization.【Methods】 We designed the primers containing Hind Ⅲ and EcoR Ⅰrestriction enzyme cutting site.The primers represented full length of PDCD5 and three other different domains.Transfected the constructed plasmids to gastric cancer cell SGC-7901.Using fluorescence microscopy to detect the cellular localization of fusion protein.【Results】 The fragment has corresponding length was detected in the products of the restriction enzyme digestion.And its sequence was validated by sequencing.Expression of the fusion protein was detected by fluorescence microscope.【Conclusion】 The plasmids representing different domains of PDCD5 protein were constructed and expressed in gastric cancer cells successfully.Four different domain proteins all located on cytoplasm.

关 键 词：PDCD5 质粒构建 胃癌细胞 PEGFP-C1 

分 类 号：Q6[生物学—生物物理学] R735.2[医药卫生—肿瘤]