类硫氧还蛋白hTRXLⅠcDNA的克隆和原核表达  

Cloning and Expression of a Human Thioredoxin Like Gene ( hTRXLⅠ)

作　　者：袁红玲[1,2] 杨磊[1] 黄瑾[1] 潘泽民[1] 顾永清[1] 陈宣茂[3] 应康[3] 谢毅[3] 

机构地区：[1]石河子大学医学院 [2]复旦大学生命科学学院遗传学研究所,上海200433 [3]复旦大学生命科学学院遗传学研究所

出　　处：《复旦学报（自然科学版）》2000年第6期593-597,共5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 18周胎脑中抽提mRNA ,建立cDNA文库 ,通过大规模测序克隆出一条人类基因 ,该基因全长16 44bp ,开放阅读框 1146bp ,编码一个含 372个氨基酸的蛋白 ,预测分子质量约为 46 8ku .经与蛋白数据库比较 ,发现具有人硫氧还蛋白结构 ,命名为人的类硫氧还蛋白基因Ⅰ (humanThioredoxinlikegeneⅠ ,hTRXLⅠ ) .多组织膜Northernblot结果显示在心脏、脑、胎盘、肝、骨骼肌、肾、胰腺等各个组织中均有表达 ,肺中表达不明显 .通过Unigene染色体定位 ,将hTRXLⅠ定位于 11q11,把hTRXLⅠ的读框克隆入经改造的PBV2 2 0质粒中 ,在E .coli中诱导后获得表达 .Through screening a human fetal brain (18 weeks) cDNA library, a cDNA was isolated. It's 1 644 bp in full length and ORF was 1 146 bp. The putative protein had 372 amino acids, with molecular weight about 46.8 ku. Compared with that of protein data pool, the putative protein was found to have human thioredoxin structure. So it was named human thioredoxin like gene Ⅰ ( hTRXL Ⅰ). Northern blot analysis of multiple tissues indicated hTRXL Ⅰ expressed in heart, brain, placenta, liver, skeletal muscle, kidney, pancreas and so on, but not in lung. By using Unigene, hTRXL was mapping to human chromosome 11q11. The ORF of hTRXL Ⅰ was subcloned into reconstructed PBV220 plasmid, and found expression in BL21 with IPTG inducing.

关 键 词：类硫氧还蛋白CDNA 克隆 表达 原核表达 

分 类 号：Q78[生物学—分子生物学] Q591.2