Knauf V, Yanofsky M, Montoya A, Nester E
J Bacteriol. 1984 Nov;160(2):564-8. doi: 10.1128/jb.160.2.564-568.1984.
Agrobacterium tumefaciens Ag162 induces crown gall disease on an unusually narrow range of host plants. The 231-kilobase Ti plasmid which has been shown to determine host range, was subcloned into the vector pVCK102. By comparing overlaps of cloned insets, maps were constructed for the restriction endonucleases SalI, XhoI, EcoRI, and KpnI. Plasmid incompatibility, octopine catabolism, and at least six virulence genes were localized. Plasmid incompatibility between pTiAg162 and the wide host range plasmid pTiA6 consists of two components: mutual incompatibility and the apparent ability of pTiA6 to block RK2 replication if the pTiAg162 incompatibility locus is linked to the vector pVK102. The octopine catabolism locus maps within the 30 kilobases of DNA separating the two T-DNA regions of pTiAg162. Complementation of avirulent vir mutants of pTiA6 with clones of pTiAg162 DNA did not confer the host range of pTiAg162 but rather restored the wide host range of pTiA6. One potentially important difference between pTiA6 and pTiAg162 is that pTiAg162 T-DNA regions are widely separated.
根癌土壤杆菌Ag162仅在异常狭窄的宿主植物范围内诱发冠瘿病。已证明决定宿主范围的231千碱基Ti质粒被亚克隆到载体pVCK102中。通过比较克隆片段的重叠部分,构建了限制性内切酶SalI、XhoI、EcoRI和KpnI的图谱。确定了质粒不相容性、章鱼碱分解代谢以及至少六个毒力基因的位置。pTiAg162与广宿主范围质粒pTiA6之间的质粒不相容性由两个成分组成:相互不相容性以及如果pTiAg162不相容位点与载体pVK102相连,pTiA6阻止RK2复制的明显能力。章鱼碱分解代谢位点位于pTiAg162两个T-DNA区域之间相隔30千碱基的DNA范围内。用pTiAg162 DNA克隆对pTiA6无毒力的vir突变体进行互补,并未赋予pTiAg162的宿主范围,而是恢复了pTiA6的广宿主范围。pTiA6和pTiAg162之间一个潜在的重要差异是pTiAg162的T-DNA区域相距很远。