Molbas Research Group, Department of Plant Molecular Biology, University of Leiden, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands.
EMBO J. 1984 Nov;3(11):2485-90. doi: 10.1002/j.1460-2075.1984.tb02160.x.
The intact T-region of the B6Ti plasmid of Agrobacterium tumefaciens was stepwise cloned into a site in transposon Tn3. In this way a suitable vehicle (Tn1882) was obtained for translocating the T-region to different replicons, i.e., to other plasmids or the chromosome. The IncP plasmid R772::Tn1882 conferred tumorigenicity on Agrobacterium if the virulence genes were provided in trans in the same cell. This result showed that the T-region present on Tn1882 was transferred efficiently to plant cells. Normal tumor development also occurred if the T-region was placed in the chromosome of A. tumefaciens and an R' plasmid was present carrying virA-E or virA-F. We conclude that the plasmid location of the T-region is not a prerequisite for transfer to the plant cell. The apparently normal delivery of the T-DNA from a bacterial chromosomal location supports a model involving a processing step within Agrobacterium effecting transfer of the T-region as a separate entity.
将完整的 B6Ti 质粒 T 区分步克隆到转座子 Tn3 的一个位点中。通过这种方式,获得了一个合适的载体(Tn1882),可将 T 区转移到不同的复制子,即其他质粒或染色体上。如果在同一细胞中转录毒性基因,IncP 质粒 R772::Tn1882 会使根癌农杆菌具有致瘤性。该结果表明,Tn1882 上的 T 区可有效地转移到植物细胞中。如果 T 区位于根癌农杆菌的染色体上,并且存在携带 virA-E 或 virA-F 的 R'质粒,则正常的肿瘤发育也会发生。我们得出结论,T 区的质粒位置不是转移到植物细胞的必要条件。从细菌染色体位置明显正常地传递 T-DNA,支持了一种模型,即涉及在根癌农杆菌内的一个加工步骤,将 T 区作为一个独立的实体进行转移。