整合素β4结合蛋白与P311在人胚肾293细胞中的表达及共定位  

Expression and co-localization of integrin beta 4 binding protein and P311 in human embryo kidney 293 cells

作　　者：易绍萱[1] 袁顺宗[2] 马兵[1] 贺伟峰[2] 陈希炜[2] 胡晓红[2] 张小容[2] 彭旭[2] 周丽娜[2] 罗高兴[2] 吴军[2] 

机构地区：[1]成都医学院第一附属医院实验研究中心,四川省成都市610500 [2]解放军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,重庆市市级重点实验室,重庆市400038

出　　处：《中国组织工程研究与临床康复》2007年第36期7211-7214,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金项目资助(30571922)~~

摘　　要：目的:利用激光共聚焦显微镜共定位检测整合素β4结合蛋白与P311蛋白在细胞内的分布,进一步证实它们在细胞内产生相互作用的可能性。方法:实验于2006-03/12在解放军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完成。先分别构建能在哺乳动物细胞中表达整合素β4结合蛋白的绿色荧光融合蛋白和表达P311的红色荧光融合蛋白的重组载体,经酶切鉴定正确后,利用脂质体介导法共转染人胚肾293细胞后,激光共聚焦显微镜进行共定位观察。结果:表达整合素β4结合蛋白的绿色荧光融合蛋白和表达P311的红色荧光融合蛋白的重组载体经双酶切鉴定正确,转染293细胞后,荧光显微镜下能观察到绿色和红色荧光。激光共聚焦显微镜下观察两者主要分布在293细胞的胞浆和胞核中,并且存在着共定位现象。结论:表达整合素β4结合蛋白的绿色荧光融合蛋白和表达P311的红色荧光融合蛋白的重组载体在空间分布上两者存在相互作用的物质基础。AIM： To observe the distribution of integnn beta 4 binding protein （ITGB4BP） and P311 protein in cells by confocal laser scanning microscopy and further vedfy the possibility of the interaction between ITGB4BP and P311 in cells. METHODS： The experiment was performed at the State Key Laboratory of Trauma, Bums and Combined Injury, Institute of Bums, Southwest Hospital, Third Military Medical University of Chinese PLA from March to December 2006. Target fragments P311 and ITGB4BP were subcloned into Red Fluorescent Protein （RFP） expression vector and Green Fluorescent Protein （GFP）, respectively. After the recombinants identified by restriction enzyme digestion, the two constructions were co-transfected into human embryo kidney 293 （HEK293）cells and the expression products were detected by confocal laser scanning microscopy. RESULTS： Double restriction enzyme digestion showed that the two recombinant vectors were constructed correctly. Both vectors could express in HEK293 cells, and green and red fluorescence were observed under fluorescent microscope. The fluorescence fusion proteins mainly distributed in the cytoplasm and nuclei under confocal laser scanning microscopy and were co-localized. CONCLUSION： The recombinant vectors of GFP-ITGB4BP and RFP-P311 have interactive matedal basis in spatial distribution.

关 键 词：整合素β4结合蛋白 P311 激光共聚焦 基因表达 共转染 组织构建 

分 类 号：R619.6[医药卫生—外科学]