DEVELOPMENT OF EXPRESSION CLONING SYSTEM FOR ONCOGENE CDNAS

癌基因 CDNAS 表达克隆系统的开发

• 批准号: 3874710
• 负责人: T MIKI
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3874710
• 关键词: autosomal dominant trait; autosomal recessive trait; complementary DNA; eukaryote; gene expression; gene rearrangement; genetic library; genetic manipulation; genetic promoter element; genetic recombination; human tissue; molecular cloning; neoplasm /cancer genetics; neoplastic cell; neoplastic transformation; nucleic acid sequence; oncogenes; plasmids; protooncogene; tissue /cell culture; transfection; transposon /insertion element

An efficient expression cloning system has been developed to isolate cDNAs which can change cell morphologies. A cDNA library was constructed from a transformant induced by mouse hepatocellular carcinoma DNA, and the library DNA was used to transfect NIH/3T3 cells. From two of the several transformed foci obtained, identical plasmids with transforming capability were rescued. Analysis of the cDNA clones for the oncogene, designated hep1, showed that the gene was created by recombination events between an unknown gene and mouse homologue of the B-raf proto-oncogene. This genetic rearrangement was detected in two primary transformants independently induced by the original tumor DNA, suggesting that the rearrangement generating the hep1 oncogene occurred somatically, in contrast to human B-raf oncogene which was created in vitro during DNA transfection.

一个高效的表达克隆系统已被开发出来，以分离cDNA 可以改变细胞形态构建了一个cDNA文库， 小鼠肝癌DNA诱导的凋亡及文库 DNA转染NIH/3 T3细胞。从几个中的两个 获得转化灶，具有转化能力的相同质粒 获救了分析癌基因的cDNA克隆，命名为 hep 1，表明该基因是由一个 未知基因和B-raf原癌基因的小鼠同源物。这种遗传 在两个初级转化体中独立地检测到重排 诱导的原始肿瘤DNA，这表明重排， hep 1癌基因的产生发生在体细胞，与人类相反， B-raf癌基因是在DNA转染过程中体外产生的。