大鼠小脑特异基因Zic的克隆、序列分析和初步表达  

Cloning, Sequencing and Preliminary Expression of Novel Zinc Finger Gene Expressed Restrictedly in Rat Brain

作　　者：蔡莹芸 康建胜[2] 刘海兰[2] 刘建华[1] 李荣秀[2] 

机构地区：[1]上海交通大学生命科学技术学院,上海200240 [2]中国科学院上海生命科学研究院生物工程研究中心

出　　处：《上海交通大学学报》2001年第7期1071-1075,共5页Journal of Shanghai Jiaotong University

基　　金：中国科学院生物科学与生物技术研究特别支持费课题 (财政部专项 )

摘　　要：EST(AW0 55732 )片段是 SD大鼠脑内一特异表达基因 c DNA的 3′-端片段 ,与小鼠和人的Zic基因同源 .基于小鼠及人的 Zic基因的同源序列 ,设计引物用 PCR方法从 SD大鼠脑中克隆得到该基因的编码区序列 (r Zic) .该基因编码一锌指蛋白 ,其锌指结构域与线虫的 tra- 1基因 ,果蝇的ci D基因 ,人的 Gli癌基因和果蝇的 opa基因的锌指结构域高度同源 .用 RT- PCR方法分析了 r Zic基因在大鼠脑区分布状况 .结果表明 ,r Zic基因在小脑中高表达 .将该 r Zic基因克隆至带有 6个组氨酸的表达载体 p ET- 32 a(+)中 ,用 IPTG诱导 r Zic融合蛋白在大肠杆菌 BL2 1菌株中表达 ,诱导后重组蛋白质占菌体总蛋白的 2 0 .8% .EST fragment (AW057732) is a 3′ end cDNA fragment which expresses highly restrictedly in the brain of SD rat. It shows homologous with the Zic gene of both mouse and human. Based on the homology of the sequences of mouse and human, the full length cDNA of the coding region from the rat brain (named rZic ) was got. This gene encodes a zinc finger protein, whose zinc finger domain shows high homology with Caenorhabditis elegans tra 1 gene, Drosophila ci D gene, human Gli oncogene and Drosophila opa gene. Through the method of RT PCR, we found that it expresses affluently in the cerebellum of the SD rat. Therefore, this novel gene may have some relation with the development of the cerebellum. rZic gene was cloned into the expression vector pET 32a(+). The recombinant pET 32a(+) rZic was transformed into E.coli strain BL21 and the expression of a 6 His tagged rZic fusion protein was induced by IPTG. Band density scaning of stained gel was performed to estimate the percentage of the recombinat protein in the total bacterial protein. The ratio is 20.8%.

关 键 词：Zic基因 锌指结构 小脑 SD大鼠 基因克隆 序列分析 锌指蛋白 

分 类 号：Q785[生物学—分子生物学] Q754