MOLECULAR CLONING AND NUCLEOTIDE SEQUENCE ANALYSIS OF A TRANSFORMING GENE FROM HUMAN GASTROCARCINOMA CELL LINE

Mouse and rat fibroblasts were transfected with total DNA from human gastrocarcinoma cell line BGC-823. It was shown by hybridization assay that the genome of one of the rat secondary foci contains transforming genes from the human gastrocarcinoma cell line, which are homologous to the protooncogene c-Ha-ras in the normal cells. The genomic library of the rat secondary foci was constructed, using λ phage EMBL3 as the vector. The transforming gene Ha-ras of the human gastrocarcinoma cell was thus cloned by screening the library with the probes of human Alu repeat sequence and c-Ha-ras. The nucleotide sequences of the first and second exons were analysed by M13-dideoxy method. The result shows that the nucleotide sequence of the transforming gene is the same as that of the normal protooncogene except one nucleotide difference in the first exon.     

人胃癌细胞株转化基因的克隆和核苷酸序列的分析

本文用人胃癌细胞株BGC-823总DNA对小鼠及大鼠成纤维细胞进行转染。用分子杂交方法证明大鼠第二轮恶性转化细胞基因组中合有来自人胃癌的转化基因,它与存在于正常细胞中的原癌基因c-Ha-ras同源。以λ噬菌体EMBL 3为载体,构建大鼠第二轮转化细胞基因组文库,以人Alu重复序列和c-Ha-ras为探针,将人胃癌细胞转化基因Ha-ras从文库中克隆分离出。用M13——双脱氧法测定转化基因第一、第二外显子核苷酸序列。结果表明转化基因除了第一外显子中有一个碱基发生变化外,核苷酸序列与原癌基因完全相同。