人类抗砷相关基因(hARRG)的克隆和表达  

A cloning expression of human arsenite resistance related gene(hARRG)

作　　者：杨磊[1] 王国荃[1] 应康[2] 黄瑾[3] 戴建凉[2] 袁有忠[2] 刘建平[2] 谢毅[2] 毛裕民[2] 

机构地区：[1]新疆医科大学公共卫生学院环境卫生学教研室,新疆乌鲁木齐830054 [2]复旦大学遗传所国家遗传工程重点实验室,上海200433 [3]石河子大学医学院,新疆石河子832002

出　　处：《新疆医科大学学报》2000年第2期95-97,共3页Journal of Xinjiang Medical University

摘　　要：目的 :克隆并表达人类抗砷基因 human arsenite resistance related gene(h ARRG)。 方法 :从人胎脑中获得 m RNA,建立 c DNA文库 ,通过大规模测序克隆出一人类新基因。 结果 :该基因全长 1170 bp,读框为 (ORF)72 3bp,经序列分析 ,与中国仓鼠抗砷基因 arsenite resistance gene (asr2 )同源性达 88%。根据 ORF设计引物 ,PCR扩增 ,扩增产物酶切后装入 PQE30质粒 ,诱导后获得 >40 %的高效表达。用镍亲和层析法纯化后纯蛋白回收率 >80 %。 结论 :首次在国内成功克隆并表达了人类抗砷相关基因 h ARRG。Objective: To study cloning expression of human arsenite resistance related gene(hARRG). Methods: Obtaining mRNA from human fetal brain, constructing cDNA library, then cloning a new human gene by sequencing of wide scope.Results: The gene is 1170 bp long, its opening reading frame (ORF)is 723 bp long. Through sequential analysis, its homologous amount was 88% identical to Chiness hamster arsenite resistance gene(asr2). We designed primer according to ORF, with polymerize chain reaction(PCR); the hARRG was acquired from human fetal brain DNA and was inserted into pQE30 plasmid. The highly effectively expressed protein was gained about 40% after IPTG induction. With nickel alloy layer analysis ,the rate of recycling of purified protein was more than 80%. Conclussion: hARRG was successfully cloned and expressed from human fetal brain DNA.

关 键 词：抗砷基因 克隆 砷中毒 hARRG 基因表达 

分 类 号：R595.2[医药卫生—内科学]