HT036是纤维化相关基因P311的相互作用蛋白  

HT036 as the novel interactive protein of fibrotic-related P311

作　　者：彭旭[1] 谭江琳[1] 袁顺宗[1] 贺伟峰[1] 陈烯伟[1] 罗高兴[1] 吴军[1] 

机构地区：[1]第三军医大学西南医院全军烧伤研究所创伤,烧伤与复合伤国家重点实验室,疾病蛋白质组学重庆市市级重点实验室,重庆400038

出　　处：《创伤外科杂志》2009年第5期412-415,共4页Journal of Traumatic Surgery

基　　金：国家自然科学基金面上项目(30571922;30672174);创伤烧伤复合伤国家重点实验室自主研究课题项目资助(SKLZZ200808)

摘　　要：目的利用激光共聚焦显微镜观察未知蛋白(HT036)与纤维化相关蛋白(P311)的细胞内表达与共定位。方法构建带绿色荧光的HT036真核表达载体,经鉴定正确后,与前期构建带红色荧光的P311真核表达载体共转染人胚肾上皮细胞株(HEK293),同时共转染空载体做对照,激光共聚焦显微镜下观察蛋白在细胞内表达、分布和共定位。结果HT036真核表达载体经聚合酶链反应(PCR)、双酶切和测序鉴定正确,与P311共转染HEK293细胞后,激光共聚焦显微镜观察到绿色和红色荧光表达。HT036和P311主要分布在细胞胞浆,两者在HEK293细胞中存在共定位,而对照组则无该现象。结论通过酵母双杂交筛选出的P311相互作用蛋白HT036,在活体细胞内存在空间上的共定位,进一步证明它们之间的相互作用,为探讨P311纤维化机制提供新的切入点。Objective To observe the intracellular expression and the co-location of HT036 with P311 in human embryo kidney cell line. Methods The encoding sequence of HT036 was amplified and cloned into the eukaryotic express/on plasmid pEGFP-N2. The construction was verified by sequencing and then co-transfected into HEK293 with pDsRed-P311. The expression products were visualized and analyzed by confocal microscope. Results The vector was successfully constructed. The confocal microscope scanning showed that the fluorescence fusion proteins mainly distributed in cytoplasm and there was co-location between the two proteins. Conclusion This study further confirmes the issue that I-IT036 and P311 may interact with each other in vitro and play role in hypertrophic scar formation.

关 键 词：创伤修复 蛋白 基因 成纤维细胞 瘢痕 

分 类 号：R641[医药卫生—外科学]