Bisaro D M, Hamilton W D, Coutts R H, Buck K W
Nucleic Acids Res. 1982 Aug 25;10(16):4913-22. doi: 10.1093/nar/10.16.4913.
We report the molecular cloning of the tomato golden mosaic virus (TGMV) genome in the E. coli plasmid pAT 153. The results of this work conclusively show that TGMV DNA consists of two components (designated A and B) of almost, but not exactly, the same size. Four different recombinant plasmids are described, two containing component A in opposite orientation and two containing component B in opposite orientation. Southern blot analysis revealed little sequence homology between A and B and showed both components to be equally represented in viral and intracellular DNA forms. Detailed restriction maps of the cloned DNAs are presented, and a comparison of these with digests of intracellular viral dsDNA indicates that the former are full-length faithful copies of TGMV DNA. This is the first report of the cloning of a geminivirus genome.
我们报道了番茄金色花叶病毒(TGMV)基因组在大肠杆菌质粒pAT 153中的分子克隆。这项工作的结果确凿地表明,TGMV DNA由两个大小几乎相同但并不完全相同的组分(分别命名为A和B)组成。描述了四种不同的重组质粒,其中两种以相反方向包含组分A,另外两种以相反方向包含组分B。Southern印迹分析显示A和B之间几乎没有序列同源性,并且表明这两个组分在病毒和细胞内DNA形式中均有同等程度的呈现。给出了克隆DNA的详细限制性图谱,将这些图谱与细胞内病毒双链DNA的酶切产物进行比较表明,前者是TGMV DNA的全长忠实拷贝。这是双生病毒基因组克隆的首次报道。