Coffin J M, Hageman T C, Maxam A M, Haseltine W A
Cell. 1978 Apr;13(4):761-73. doi: 10.1016/0092-8674(78)90226-x.
The genome of the Moloney strain of murine leukemia virus (Mo-MuLV) has been analyzed by digestion with ribonuclease T1 and separation of the digestion products by two-dimensional gel electrophoresis. Thirty large oligonucleotides isolated from such a fingerprint have been characterized. One of these oligonucleotides (number 21) was found to be present in twice the molar yield of the rest. The 30 oligonucleotides were mapped on the genome by determining their yields in various size classes of 3' terminal fragments of Mo-MuLV RNA. The physical map obtained in this way suggested that oligonucletoide 21 was present very near the 3' end of the geome as well as in another location near or at the 5' end. The genome structure suggested by these results was confirmed by analyzing oligonucleotides in Mo-Mulv RNA complementary to strong stop DNA, which is shown to be a copy of the 5' terminal 134 nucleotides of the MoMuLV genome. Some of the oligonucleotides in the RNA protected from RNAase digestion by hybridization to this DNA, including oligonucleotide 21, were present near both the 3' and 5' ends. Comparison of these with the nucleotide sequence of strong stop DNA shows that there is a terminal redundancy of 49-60 nucleotides in the Mo-MuLV genome RNA.
用核糖核酸酶T1消化莫洛尼氏鼠白血病病毒(Mo-MuLV)的莫洛尼株基因组,并通过二维凝胶电泳分离消化产物,以此对该基因组进行了分析。从这样一个指纹图谱中分离出的30个大的寡核苷酸已得到鉴定。发现其中一个寡核苷酸(编号21)的摩尔产量是其余寡核苷酸的两倍。通过测定它们在Mo-MuLV RNA 3'末端片段的各种大小类别的产量,将这30个寡核苷酸定位在基因组上。以这种方式获得的物理图谱表明,寡核苷酸21非常靠近基因组的3'末端,同时也位于5'末端附近或5'末端的另一个位置。通过分析与强终止DNA互补的Mo-Mulv RNA中的寡核苷酸,证实了这些结果所提示的基因组结构,强终止DNA被证明是MoMuLV基因组5'末端134个核苷酸的拷贝。通过与该DNA杂交而免受RNA酶消化的RNA中的一些寡核苷酸,包括寡核苷酸21,在3'和5'末端附近均有存在。将这些寡核苷酸与强终止DNA的核苷酸序列进行比较表明,Mo-MuLV基因组RNA中存在49 - 60个核苷酸的末端冗余。