网上实验室克隆与鉴定食管癌相关基因ECRG-4  

Using Lab On-line to Clone and Identify the Esophageal Cancer Related Gene 4

作　　者：毕美霞[1] 韩卫东[2] 陆士新[1] 

机构地区：[1]中国医学科学院 [2]中国人民解放军总医院基础医学研究所分子生物室,北京100853

出　　处：《生物化学与生物物理学报》2001年第3期259-261,共3页

基　　金：国家重点基础研究发展规划 (973)资助项目!No .G19980 5 12 0 4&&

摘　　要：利用生物信息学 ,探索网上克隆基因与鉴定基因的新方法。以因特网为平台 ,数据库为试验材料 ,各种软件为工具组成网上实验室 ,是人类基因组计划带来的实验技术革命。利用网上实验室以食管癌相关基因E CRG 4的 97bpEST为基础 ,成功克隆并鉴定了该基因。结果显示ECRG 4近似cDNA全长序列为 772bp ,其中含有一 44 7bp的完整阅读框 ,编码 148个氨基酸。氨基酸序列相似性分析表明ECRG 4与细胞膜上的免疫球蛋白超家族具有 31%同源性。该基因定位于染色体 2 q14.1～ 14.3。组织分布表明ECRG 4在正常食管、膀胱组织中表达明显高于相应的癌组织。采用辐射杂交细胞系GeneBridge 4RH嵌板作染色体定位进行初步验证 ,所得结果与网上克隆完全一致。研究提示 ,利用网上实验室克隆鉴定基因 ,是一种简便、精确的好方法。ECRGUsing Internet as platform, databases as materials and software as tools to assemble a lab on line is revolutionizing in bioscience research. The major works of lab on line are cloning, identification, localization of genes, and the structural and functional analysis of proteins. In this report, the esophageal cancer related gene 4( ECRG 4 )(accession number: AF325503) was successfully isolated. The 97 bp ECRG 4 EST was initially used to fish the human EST databases. Five pieces of ESTs showed strong homology to it, and they were assembled to one 772 bp cDNA sequence by DNASTAR software. Then the 447 bp full open reading frame of ECRG 4 was determined by ORF FINDER to encode 148 amino acids. Sequence of ECRG 4 did not reveal remarkable similarity to the known sequences in a homology analysis with the public database of GenBank, showing that it is a new gene. Homology analysis of protein coding by ECRG 4 showed a 31% homology with mouse IgG V region. ECRG 4 gene is expressed in normal esophagus, bladder and brain tissues, but its expression was significantly down regulated in prostate tumors and tumor cell lines. ECRG 4 gene was located in 2q14.1—14.3 by HTGS and STS, and was conformed by radiation hybrid (RH) method. We propose that this purely lab on line cloning approach can be used by modestly sized laboratories to rapidly and accurately characterize human genes without wasting too much money and time.

关 键 词：网上实验室 基因克隆 ECRG-4 食管癌相关基因 

分 类 号：Q78[生物学—分子生物学]