High levels of double-stranded transferred DNA (T-DNA) processing from an intact nopaline Ti plasmid.
High levels of double-stranded transferred DNA (T-DNA) processing from an intact nopaline Ti plasmid.
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来自完整胭脂碱 Ti 质粒的高水平双链转移 DNA (T-DNA) 加工。
DOI：
10.1073/pnas.86.7.2133
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发表时间：
1989
影响因子：
11.1
通讯作者：
Kado,CI
中科院分区：
文献类型：
--
作者：
Steck,TR;Close,TJ;Kado,CI
为了获得植物的细菌介导的致癌转化，在感染期间将根癌农杆菌的肿瘤诱导（Ti）质粒的转移DNA（T-DNA）转移到其植物宿主细胞中。转化的初始阶段涉及在诱导位于Ti质粒上的vir基因后在细菌细胞中加工T-DNA。检测了这种加工的动力学和条件，并且在用乙酰丁香酮诱导后，高达40%的T-DNA的左右边界被切割。这种切割依赖于virA、virG和VirD，并且不依赖于rec-independent。在诱导后30分钟内观察到加工的T-DNA，并且通过诱导培养基中磷酸盐浓度的增加而延迟。当在不存在蛋白酶处理的情况下分离DNA时，在左右边界区域处对应于切口左侧的DNA片段表现出凝胶阻滞，这表明一个或多个“先导”蛋白可能参与T-DNA转移。虽然加工产物的相对丰度并不一定意味着相对重要性，但双链切割产物的优势表明双链T-DNA应被视为T-DNA转移的可能中间体。
To obtain bacterial-mediated oncogenic transformation of plants, the transferred DNA (T-DNA) of the tumor-inducing (Ti) plasmid of Agrobacterium tumefaciens is transferred to its plant host cells during infection. The initial phases of transformation involve the processing of the T-DNA in the bacterial cell after induction of the vir genes located on the Ti plasmid. The kinetics and conditions of this processing were examined and upon induction with acetosyringone up to 40% of the left and right borders of the T-DNA were cleaved. This cleavage was dependent upon virA, virG, and VirD and was rec-independent. Processed T-DNA was observed within 30 min after induction and was delayed by an increased concentration of phosphate in the induction medium. When DNA was isolated in the absence of protease treatment, the DNA fragment corresponding to the left side of the cut at both the left and right border region exhibited gel retardation, suggesting one or more "pilot" proteins may be involved in T-DNA transfer. Although the relative abundance of a processed product does not necessarily imply relative importance, the preponderance of double-stranded cleavage products suggests that double-stranded T-DNA should be considered as a possible intermediate in T-DNA transfer.