砷代谢相关的全长新cDNA的克隆和功能初步研究  

Cloning and Preliminary Functional Analysis of a Novel Human Full-length cDNA Related to Gene Arsenite Metabolism

作　　者：顾永清[1,2] 杨磊[1] 袁红玲[1] 吴超群[3] 潘泽民[1] 夏放[3] 李瑶[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603-608,共6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：从人胎脑中获得mRNA建立cDNA文库 ,通过大规模测序克隆出一人类新基因cDNA .全长 2 2 5 5bp ,开放阅读框 (ORF) 15 32bp .用 4μmol/L砷刺激L 0 2细胞 2周 ,与未用砷刺激的L 0 2细胞作表达谱基因芯片杂交 ,发现该基因在砷刺激L 0 2细胞中表达量上调 3倍 .用不同浓度砷染毒L 0 2细胞 2周 ,与未用砷染毒的L 0 2细胞抽RNA转膜作Northern杂交 ,结果显示该基因在未染毒L 0 2细胞中几乎不表达 ;而在砷染毒细胞中该基因的表达量明显增加 ,且有随剂量增加表达量亦增加的趋势 ;同时Northern结果显示在 2 3kb处有单一条带 .用不同浓度砷染毒L 0 2细胞 2周 ,作细胞原位杂交 ,结果显示该基因在未染毒细胞中几乎不表达 ,在各染毒组细胞中均有表达 .据此 ,认为这可能是一条新的全长砷代谢相关基因 ,经HUGO/GDB人类基因命名委员会的同意 ,命名为ARG1(arseniterelatedgene 1) .After large scale sequencing analysis of a human fetal brain cDNA library constructed by our laboratory, a cDNA clone with a complete coding sequence was isolated. The clone was 2 255 bp long and contained an ORF of 1 532 bp encoding a 510 amino acids with putative molecular weight of 58.9 ku. Comparing the hybridization patterns of cDNA microarray of cultured cell (l 02 cell) with and without high dosage of arsenite induction indicated that the expression level of the cloned cDNA was elevated in arsenite induced cells. This result was confirmed by northern blot analysis, i.e, the gene expression level was higher in arsenite induced cell and increased proportionally with the arsenite dosage. In situ hybridization of L 02 cells treated with different dosages of arsenite and non treated showed that the gene expression was very low in non treated cells and higher in arsenite treated cells with a dose reaction relationship. So the gene cloned was a novel arsenite related gene and was named ARG 1 (arsenite related genel) with the agreement of HUGO/GDB.

关 键 词：砷代谢相关基因 L-02细胞 基因芯片杂交 人 抗砷基因 砷中毒 

分 类 号：R595[医药卫生—内科学] Q78[医药卫生—临床医学]