ISOLATION OF NOVEL ONCOGENES BY AN EFFICIENT EXPRESSION CLONING SYSTEM

通过高效表达克隆系统分离新型癌基因

• 批准号: 3752681
• 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/3752681
• 关键词: 3T3 cells; Baculoviridae; animal genetic material tag; athymic mouse; biological signal transduction; complementary DNA; cytoplasm; gene expression; genetic library; guanine nucleotides; molecular cloning; neoplastic transformation; oncogenes; oncoproteins; osteosarcoma; phosphorylation; serine

Using an efficient expression cloning system developed in the lab. a novel oncogene. designated ost was isolated from a rat osteosarcoma cDNA library. The cloned ost cDNA possesses high transforming activity in NIH/3T3 cells. The ost product was activated by truncation of its N- terminal domain and was highly tumorigenic in nude mouse assays. The full-length ost cDNA was subsequently isolated and encodes a predicted protein of 100 kilodaltons containing the OH (dbl homology) and PH (pleckstrin homology) domains. Ost is mainly phosphorylated on serine and localized in the cytoplasm. Among the tissues examined. brain showed the highest expression of Ost. especially in neurons and alpha-tanycytes. Baculovirus-expressed Ost can catalyze guanine nucleotide exchange on RhoA and Cdc42 among Rho and Ras family members tested. Ost did not detectably associate with RhoA or Cdc42. but interacted specifically with the GTP-bound form of Rac1. These results implicate Ost as a critical regulatory component which links the signal transduction pathways that flow through Rac1, RhoA and Cdc42.

使用实验室开发的高效表达克隆系统。一个