人Rab蛋白cDNA的克隆和表达  

Cloning, Sequencing and Expression of a Novel cDNA Coding for Human Homolog of Rab Protein

作　　者：陈宣茂[1] 孙朝辉 王兆[1] 杨泉胜[2] 应康[1] 谢毅[1] 毛裕民[1] 

机构地区：[1]复旦大学生命科学学院,遗传学研究所上海200433 [2]浙江大学生物系,浙江杭州310027

出　　处：《复旦学报（自然科学版）》2000年第6期688-691,共4页Journal of Fudan University：Natural Science

基　　金：上海市现代生物与新药产业发展基金资助项目!(98431912 1)

摘　　要：从人胎脑cDNA文库中克隆到一种新的RabcDNA ,全长 92 0bp ,拟编码 2 13个氨基酸残基 ,该蛋白预测的分子质量为 2 45 6 7u ,等电点 7.34,经同源比较 ,该cDNA与GenBank数据库中登录号为X1496 4的Rab蛋白有 83 %的相似性和 76 %的相同性 .将该cDNA克隆到经改造的PBV2 2 0表达质粒 ,转化DH5α菌株诱导表达出该蛋白 .取 2 4种不同组织的总cDNA各 10 0ng ,用该基因序列设计引物作PCR ,结果在胎肝组织中检测到有明显条带 ,表明该Rab基因相对在胎肝有高表达 .A cDNA clone coding for human Rab protein has been isolated by screening human 18 weeks embryo brain cDNA library and sequenced. This new cDNA clone of 920 bp contains an open reading frame of 213 amino acids whose deduced molecule weight is 24 567 u and isoelectronic point is 7.34. The entire amino acid sequence of novel human Rab was 83% similar to that of Rab protein Q14964 in GenBank and their sequence in the GTP binding domain were almost completely identical, predicting that the novel cDNA is for a human Rab isoform. Expression profile analysis has indicated that the human novel Rab gene was high expressed in fatal liver tissue. This novel human Rab gene was expressed in vitro using modified PBV220 expression vector.

关 键 词：RAB基因 CDNA克隆 原核表达 表达谱分析 人类 克隆 表达 

分 类 号：Q987[生物学—遗传学] Q591.2[生物学—人类学]