人类组织中TRAF3IP3基因表达的检测及转染胚肾HEK293细胞后的亚细胞定位  

Expression of human TRAF3IP3 gene in human different organs and its subcellular localization in transfected HEK 293 cells

作　　者：王洋[1] 王沂[1] 余源[1] 刘青[1] 毛焕 熊艳杰 张秀军[1] 

机构地区：[1]河北联合大学生命科学学院,河北唐山063000 [2]附属医院病理科

出　　处：《第三军医大学学报》2013年第14期1447-1450,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(81072093)~~

摘　　要：目的克隆TRAF3IP3基因,明确TRAF3IP3在部分人类组织中的表达谱并鉴定其表达产物的亚细胞定位。方法采用实时定量PCR检测TRAF3IP3基因在人体各组织中的表达情况;克隆TRAF3IP3基因开放读码框区(ORF)并构建真核亚细胞定位质粒。转染人胚肾HEK 293细胞,激光共聚焦显微镜观察融合蛋白的亚细胞定位。结果实时定量PCR证实TRAF3IP3基因在被检测的11种组织中均有表达,其中在淋巴结、脾、骨髓、胃和睾丸中的表达水平较高;成功克隆了TRAF3IP3基因ORF区并构建成为荧光定位质粒,激光共聚焦显微镜观察证实TRAF3IP3蛋白主要定位于核膜,在核周呈颗粒状分布。结论 TRAF3IP3基因在淋巴细胞富集的器官呈高表达,可能参与免疫系统的发育、成熟及调控,其表达产物在细胞内主要定位于核膜及核周胞浆。Objective To determine the tissue expression profile of human TRAF3IP3 gene, also known as TRAF3-interacting Jun N-terminal kinase-activating modulator （T3JAM） and its subcellular localiza- tion. Methods The mRNA expression of TRAF3IP3 was detected in 11 human tissue samples, including the liver, thyroid, colon, spleen, stomach, testis, uterus, fat tissue, bone marrow, lung and lymph node by real- time PCR. The subcellular location plasmid containing the fuU-length TRAF3IP3 cDNA was constructed. And then the recombinant plasmid was transfected into HEK 293 cells. Subcellular location of TRAF3IP3 was detec- ted by confocal laser scanning microscopy. Results TRAF3IP3 mRNA was expressed in all detected human tissues, and highly expressed in the lymph node, spleen, bone marrow, stomach and testis. The human TRAF3IP3 eukaryotic expression vector was constructed successfully. Confocal laser scanning microscopy showed TRAF3IP3 protein was located in the nuclear membrane, in a perinuclear granular distribution. Conclusion TRAF3IP3 mRNA is highly expressed in the lymphocyte-richened tissues, and TRAF3IP3 protein is mainly located in the nuclear membrane and perinuclear region, which might be involved in the development, maturation and regulation of immune system.

关 键 词：TRAF3IP3 组织表达谱 亚细胞定位 

分 类 号：R322.61[医药卫生—人体解剖和组织胚胎学] R394.2[医药卫生—基础医学]