Shields A, Goff S, Paskind M, Otto G, Baltimore D
Cell. 1979 Dec;18(4):955-62. doi: 10.1016/0092-8674(79)90208-3.
Virions produced from cells transformed by A-MuLV contain a 30S, 5.6 kb RNA that can be translated in a cell-free system to form the characteristic A-MuLV protein. This RNA was mapped by heteroduplex methods using DNA probes from M-MuLV, the presumed parent of A-MuLV. The overall organization of the RNA was determined by using full-length M-MuLV reverse transcribed DNA and visualizing the heteroduplexes in the electron microscope. This showed that A-MuLV and M-MuLV have homologous sequences at both ends of their RNAs but that the central portion of the A-MuLV genome is not homologous to sequences in M-MuLV RNA. A precise measure of the lengths of the shared regions was obtained by using S1 nuclease to digest hybrids between 32P-labeled M-MuLV DNA and A-MuLV RNA; the resulting fragments were analyzed for their length by electrophoresis. The regions of homology were shown to be 1320 nucleotides long at the 5' end and 730 nucleotides long at the 3' end. Thus approximately 6200 nucleotides of the approximately 8300 in M-MuLV RNA were deleted when the A-MuLV genome was formed, but an insert of 3600 nucleotides, presumably derived from the normal murine genome, was inserted in place of the deleted region.
由A-MuLV转化的细胞产生的病毒粒子含有一种30S、5.6 kb的RNA,它可以在无细胞系统中进行翻译,以形成特征性的A-MuLV蛋白。使用来自M-MuLV(A-MuLV的推测亲本)的DNA探针,通过异源双链方法对这种RNA进行了定位。通过使用全长M-MuLV逆转录DNA并在电子显微镜下观察异源双链体,确定了RNA的整体结构。这表明A-MuLV和M-MuLV在其RNA的两端具有同源序列,但A-MuLV基因组的中央部分与M-MuLV RNA中的序列不同源。通过使用S1核酸酶消化32P标记的M-MuLV DNA和A-MuLV RNA之间的杂交体,获得了共享区域长度的精确测量值;通过电泳分析所得片段的长度。同源区域在5'端显示为1320个核苷酸长,在3'端为730个核苷酸长。因此,在形成A-MuLV基因组时,M-MuLV RNA中约8300个核苷酸中的约6200个被删除,但插入了一个大概源自正常小鼠基因组的3600个核苷酸的片段来取代被删除的区域。