Canaani E, Robbins K C, Aaronson S A
Nature. 1979 Nov 22;282(5737):378-83. doi: 10.1038/282378a0.
A cleavage map of the Moloney murine sarcoma viral DNA was constructed and compared with that of a spontaneously occurring deletion mutant. By restriction enzyme analysis, it was shown that a region encompassing over 40% of the viral information was not essential for transformation or rescue of the deletion mutant. The transforming region was further localised by analysis of the transforming activity in tissue culture of isolated restriction fragments of linear duoble-stranded sarcoma viral DNA. In each case, DNA fragments that retained transforming activity preserved the cell-derived insertion sequences of the viral genome. Moreover, such transformants invariably expressed RNA specific to this region. By these two approaches, it was possible to demonstrate that the transforming region of the viral genome begins very near or within the cell-derived insertion sequences. Thus, the transforming gene of this mammalian sarcoma virus originates from within the mouse cell genome.
构建了莫洛尼鼠肉瘤病毒DNA的切割图谱,并与自发出现的缺失突变体的图谱进行了比较。通过限制性内切酶分析表明,一个包含超过40%病毒信息的区域对于缺失突变体的转化或拯救并非必需。通过分析线性双链肉瘤病毒DNA的分离限制性片段在组织培养中的转化活性,进一步定位了转化区域。在每种情况下,保留转化活性的DNA片段保留了病毒基因组的细胞衍生插入序列。此外,此类转化体总是表达该区域特有的RNA。通过这两种方法,有可能证明病毒基因组的转化区域始于非常接近细胞衍生插入序列或在其内部。因此,这种哺乳动物肉瘤病毒的转化基因起源于小鼠细胞基因组内。