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Expression cDNA cloning of a transforming gene encoding the wild-type G alpha 12 gene product.

机译:编码野生型G alpha 12基因产物的转化基因的表达cDNA克隆。

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摘要

Using an expression cDNA cloning approach, we examined human tumor cell lines for novel oncogenes that might evade detection by conventional techniques. We isolated a transforming sequence that was highly efficient in transforming NIH 3T3 mouse fibroblasts. DNA sequence analysis identified the gene as the human homolog of a recently cloned alpha subunit of mouse GTP-binding protein G alpha 12. NIH 3T3 cells transfected with G alpha 12 cDNA grew in soft agar and were tumorigenic in nude mice. There were no apparent mutations in the cloned cDNA in comparison with a G alpha 12 cDNA clone isolated from a normal human epithelial cell library, implying that overexpression alone was sufficient to cause NIH 3T3 cell transformation. The observed altered growth properties mediated by G alpha 12 showed a certain degree of dependency on serum factors, and its mitogenic potential was also potently inhibited by suramin treatment.
机译:使用表达cDNA克隆方法,我们检查了人类肿瘤细胞系中可能逃避常规技术检测的新型致癌基因。我们分离出了一种高效转化NIH 3T3小鼠成纤维细胞的转化序列。 DNA序列分析将该基因鉴定为最近克隆的小鼠GTP结合蛋白G alpha 12的alpha亚基的人类同源物。转染了G alpha 12 cDNA的NIH 3T3细胞在软琼脂中生长,在裸鼠中具有致瘤性。与从正常人上皮细胞文库中分离的G alpha 12 cDNA克隆相比,克隆的cDNA中没有明显的突变,这表明单独的过表达足以引起NIH 3T3细胞转化。观察到的由G alpha 12介导的改变的生长特性显示出对血清因子的一定程度的依赖性,并且苏拉明处理也有效抑制了它的促有丝分裂潜力。

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