移行上皮反应基因TERE1的结构与功能分析  

Structural and functional analysis of transitional epithelial response gene TERE1

作　　者：夏炎枝[1] 赵为[1] 魏雄[1] 吴时敏[2] 王西明[3] 红凌[1] 

机构地区：[1]华中科技大学生命科学与技术学院分子生物物理教育部重点实验室,湖北武汉430074 [2]华中科技大学同济医学院附属同济医院病理研究所病理学与病理生理学教育部国家重点学科,湖北武汉430030 [3]华中科技大学同济医学院基础医学院生物化学与分子生物学系,湖北武汉430030

出　　处：《中国现代医学杂志》2011年第8期913-918,923,共7页China Journal of Modern Medicine

基　　金：国家自然科学基金(No:30971608);湖北省自然科学基金(No:2009CDB074);华中科技大学校自主创新基金(No:M2009050);2008教育部优秀博士学位论文创新基金资助

摘　　要：目的用生物信息软件分析移行上皮反应基因TERE1蛋白的跨膜结构,并预测分析其重要蛋白结构域功能。检测TERE1在细胞中表达定位。方法采用不同生物信息学软件预测分析TERE1的蛋白跨膜结构及重要结构域功能。采用分子克隆方法构建绿色荧光蛋白-移行上皮反应基因(pEGFP-TERE1)重组质粒,转染人膀胱癌细胞系T24,激光共聚焦显微镜观察TERE1在细胞中表达定位。结果 TERE1蛋白为多次跨膜的蛋白,保守的区域形成3个环loop 1、loop 2和loop 3;N端和loop 1含有高度保守的位点;以loop 2和loop 3之间的区域采用PROSITE软件分析显示UbiA类异戊烯转移酶家族具有一致性序列。采用菌液PCR、双酶切鉴定和DNA测序验证pEGFP-TERE1重组质粒构建成功。TERE1主要在T24细胞浆中表达。结论 TERE1为多次跨膜的蛋白,保守的区域形成3个环,UbiA类异戊烯转移酶家族具有一致性序列。TERE1主要在T24细胞浆中表达。【Objective】 To analyze the membrane protein structure of transitional epithelial response gene（TERE1） and predict its important function of protein domains by bioinformatic software.Cell localization of TERE1 was detected using Confocal.【Methods】 Using different bioinformatic software to analyze TERE1 transmembrane protein structure and its important functional domains.The recombinant plasmid pEGFP-TERE1 was constructed,then pEGFP-TERE1 vector was transformed into the bladder cancer cell line T24,the cell localization of TERE1 was observed by laser confocal microscope.【Results】 TERE1 was putative multiple transmembrane protein,the protein structure was conserved in several species,the conserved regions were formed the first loop（loop 1）,the second loop（loop 2） and the third loop（loop 3）.Loop 1 and the N terminal have several highly conserved sites through spatial structure analysis.The consistent sequences model of UbiA class isopentenyl transferase family between loop 2 and loop 3 region was： N-x（3）——x（2）-[LIMFYT]-D-x（2）——x-R——x（2）-R-x（4）-（X,any amino acid）.2.Recombinant plasmid pEGFP-TERE1 was constructed successfully through tested by bacteria PCR,double enzymes digestion and DNA sequencing,the mutations and the reading frame shift were not found.TERE1 was expressed mainly in the cytoplasm in transfect with recombinant plasmid pEGFP-TERE1.【Conclusions】 TERE1 was putative multiple transmembrane proteins,the most conservative regions were loop 1,loop 2 and loop 3,UbiA class prenyl transferase family members have consistent sequence.The eukaryotic expression plasmid pEGFP-TERE1 was successfully constructed.Confocal laser showed that TERE1 was mainly expressed in the cytoplasm.

关 键 词：移行上皮反应基因 膀胱癌 UbiA类异戊烯转移酶功能域1 生物信息 

分 类 号：R73[医药卫生—肿瘤] Q7[医药卫生—临床医学]