CHARACTERIZATION AND SUBCLONING OF A NEW HUMAN ONCOGENE

新人类癌基因的表征和亚克隆

• 批准号: 3963551
• 负责人: G VECCHIO
• 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/3963551
• 关键词: B lymphocyte; clone cells; gene expression; genetic library; genetic mapping; human tissue; lymphoma; lymphosarcoma; messenger RNA; molecular oncology; nucleic acid sequence; oncogenes

Subgenomic clones previously obtained from a cosmid clone of a new human oncogene, designated dbl for diffuse B cell lymphoma, have been utilized in order to detect the expression of this newly isolated oncogene in fresh human tumors and tumor cell lines. For this screening, a cDNA clone (representing an incomplete copy of the dbl message) obtained from a cDNA library constructed from NIH/3T3 cells in RNAs transfected with the dbl clone, has also been used. Among about 100 RNA preparations screened, positive hybridization signals have been detected by northern blotting with 10 different preparations of either fresh tumors or cell lines. The positive samples included cell lines of clear mesodermal origin, such as fibrosarcomas, rhabdo-and leio-myosarcomas. Cell lines and fresh tumor samples derived from Ewing's sarcoma (a type of tumor whose cell of origin is still controversial) were consistently found to be positive. Further characterization of the oncogene has included the screening of the cDNA library obtained from transfected NIH/3T3 cells in order to isolate the 5' missing portions of the dbl gene. Fragments of cDNA coming from this region of the gene have been obtained and will eventually lead to the construction of a full length cDNA clone.

以前从一个新人类的粘粒克隆中获得的亚基因组克隆 对于弥漫性B细胞淋巴瘤命名为db 1的癌基因已经被用于 为了检测这个新分离的癌基因在新鲜的组织中的表达， 人肿瘤和肿瘤细胞系。 对于该筛选，cDNA克隆 （代表dbl信息的不完整拷贝） 从用dbl转染的RNA中的NIH/3 T3细胞构建的文库 克隆，也被使用。 在筛选的约100种RNA制备物中， 通过北方印迹检测到阳性杂交信号， 新鲜肿瘤或细胞系的10种不同制备物。 的 阳性样品包括透明中胚层来源的细胞系，如 纤维肉瘤、横纹肌肉瘤和平滑肌肉瘤。 细胞系和新鲜肿瘤 来源于尤文氏肉瘤（一种肿瘤，其细胞来源于 仍然存在争议）一直被发现是积极的。 进一步 癌基因的特征包括筛选cDNA 从转染的NIH/3 T3细胞中获得的文库，以分离5'端 DBL基因的缺失部分。 来自于此的cDNA片段 该基因的区域已经获得，并将最终导致 全长cDNA克隆的构建。