TRANSDUCTION OF GENETIC INFORMATION RELATED TO CANCER

与癌症相关的遗传信息的转导

• 批准号: 3170034
• 负责人: PAUL BERG
• 金额: $ 17.69万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-04-01 至 1987-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3170034
• 关键词: neoplasm /cancer genetics

The identification of cellular genes whose expression or overexpression can cause cellular transformation and cancer is an important goal. The proposed researches, summarized below, are directed towards that objective. 1. A new method has been developed for cloning cellular mRNAs as cDNAs in E. coli or cultured animal cells. Since the procedure can generate nearly full length cDNA clones (the cDNA segments extend from the nucleotides adjacent to the poly A site up to, or a few nucleotides short of, the cap site) with high efficiency (about 10 5 cDNA recombinants per Mug of cell mRNA), the cloning vector has been constructed in such a way as to permit the expression of the cloned cDNA segment. Thus, it is feasible to attempt to clone cDNAs that could alter the phenotype of selected recipient cells or produce a product which permits the selection or identification of transduced cells. One objective is, therefore, to isolate cDNA copies of known and as yet unrecognized oncogenes by their ability to transduce normal to transformed cells. Similarly, cDNAs prepared from normal cells will be tested to determine if any can suppress the transformed cell phenotype of induced or natural cancer cells. 2. Studies of the structure and function of the SV40 virus early region promoter have shown that a 72 base pair tandem repeat sequence at about position -100 to -25 from the CAP-sites of the early mRNA, has a unique capacity to augment the transcribing activity of weak promoter sequences. The entire SV40 early promoter and its constituent parts (the promoter per se and the 72 bp tandem repeat) have been introduced into bacterial plasmids and will be tested for their ability to transduce normal to transfored cells by integrating at or near cellular genes whose overexpression lead to transformation. If transformed cells arise, the modified and corresponding normal genomic regions will be cloned and an analysis of their structure and oncogenic properties undertaken.

鉴定其表达或过度表达的细胞基因 可以引起细胞转化和癌症是一个重要目标。 这 拟议的研究（总结如下）旨在 客观的。 1. 开发了一种克隆细胞mRNA的新方法 作为大肠杆菌或培养的动物细胞中的 cDNA。 由于程序可以 生成几乎全长的 cDNA 克隆（cDNA 片段从 与多聚腺苷酸位点相邻的核苷酸最多或短几个核苷酸 的帽位点）具有高效率（每个约 10 5 cDNA 重组体） 一杯细胞mRNA），克隆载体是这样构建的 以允许克隆的cDNA片段的表达。 因此，它是 尝试克隆可以改变表型的 cDNA 是可行的 选择受体细胞或产生允许选择的产品 或转导细胞的鉴定。 因此，目标之一是 分离已知和尚未识别的癌基因的 cDNA 拷贝 将正常细胞转导为转化细胞的能力。 同样，cDNA 将测试由正常细胞制备的细胞是否可以抑制 诱导或天然癌细胞的转化细胞表型。 2. SV40病毒早期区结构与功能的研究 启动子显示，大约 72 碱基对的串联重复序列 早期 mRNA CAP 位点的 -100 至 -25 位，具有独特的 增强弱启动子序列转录活性的能力。 整个SV40早期启动子及其组成部分（每个启动子 se 和 72 bp 串联重复序列）已被引入细菌中 质粒并将测试它们将正常转导为 通过整合细胞基因或附近的细胞基因来转化细胞 过度表达会导致转化。 如果转化细胞出现， 修改后的和相应的正常基因组区域将被克隆，并且 对其结构和致癌特性进行分析。