重组Endocan转染MDCK细胞对其表达骨桥蛋白的影响  

The transfection of recombinant Endocan to MDCK cells and influence on OPN

作　　者：陈孝宇[1] 胡超杰[1] 左莉[1] 刘超[1] 张素梅[1] 胡若磊[1] 朱华庆[1] 周青[1] 桂淑玉[2] 汪渊[1] 

机构地区：[1]安徽医科大学分子生物学实验室、生物化学教研室和安徽省／省部共建教育部重要遗传病基因资源利用重点实验室,合肥230032 [2]安徽医科大学第一附属医院呼吸内科,合肥230022

出　　处：《安徽医科大学学报》2008年第4期381-384,共4页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金项目(编号:050430705,01043716)

摘　　要：目的构建Endocan真核表达载体pcDNA3.1-En-docan,转染犬肾小管上皮细胞MDCK,观察其在MDCK中的表达并初步分析其对骨桥蛋白(OPN)表达的影响。方法应用PCR从pET28-Endocan中扩增出Endocan cDNA全长序列,酶切后导入pcDNA 3.1/myc-his A多克隆位点,构建真核表达载体pcDNA3.1-Endocan;瞬时转染MDCK细胞,Western blot检测Endocan和OPN表达。结果经双酶切鉴定及测序分析表明Endocan已经克隆到pcDNA 3.1/myc-his A中。Western blot结果表明转染成功,Endocan表达量增加,且OPN随其而增加。结论Endocan可促进MDCK表达OPN。Objective Eukaryotic expressing plasmid pcDNA3.1-Endocan was constructed and transfected into MDCK, to observe the expression of Endocan and analyse its influence on OPN. Methods Endocan full length cDNA was amplified using PCR from pet28-Endocan, subcloned into pcDNA3.1/myc-his A, transiently transfected into MDCK cells. Western blot was used to detect the expression of Endocan and OPN. Results The construction of pcDNA3.1-Endocan had been successfully established by double enzyme digesting and sequencing. The trasfected cells expressed more Endocan and OPN. Conclusion MDCK cells express more OPN by promoting Endocan.

关 键 词：转染 基因表达 肾肿瘤/遗传学 

分 类 号：R343.9[医药卫生—基础医学] R394.2