Ti-质粒及植物基因组的相互作用.pptVIP

Ti-质粒与植物基因组的相互作用 蒋舜媛 董霞 任昶 2002-10-08 What is Ti-plasmid? Ti-plasmid, ——（short for tumor-inducing plasmid）is a large (150~200 kb) double-stranded circular DNA plasmid in a soil bacterium called Agrobacterium tumefaciens, which causes what is known as produces uncontrolled growths (tumors, or galls), normally at the base (crown) of the plant. Ti-plasmid is the key to this tumor production. Figure 1. In the process of causing crown gall disease, the bacterium Agrobacterium tumefaciens inserts a part of its Ti plasmid—a region called T-DNA—into a chromosome of the host plant. Figure 2. Simplified representation of the major regions of the Ti plasmid of A. tumefaciens. The T-DNA, when inserted into the chromosomal DNA of the host plant, directs the synthesis of nopaline, which is then utilized by the bacterium for its own purposes. T-DNA also directs the plant cell to divide in an uncontrolled manner, producing a tumor. Structure of Ti-plasmid T-DNA区 毒性区 (Vir-region, vir) 质粒复制起点 (Origin of replication, ori) 质粒结合转移位点 (Transfer function site, tra) 冠瘿碱分解位点 (ocs or nos) Two types of Ti-plasmid OCS plasmid (octopine ) pTiAch5, pTiA6NC, pTiB653, pTiAg162 NOS plasmid (nopaline ) pTiT37, pTiT38 土壤农杆菌-植物DNA转移体系步骤 植物敏感细胞和土壤农杆菌相互作用 土壤农杆菌的毒性区基因被激活 T-DNA的切割和T复合物生成 T复合物由土壤农杆菌经植物细胞膜进入植物细胞 T-DNA整合到植物染色体上 Figure 1. In the process of causing crown gall disease, the bacterium Agrobacterium tumefaciens inserts a part of its Ti plasmid—a region called T-DNA—into a chromosome of the host plant. Interaction of Agrobacterium Ti-plasmid DNA and plant cells植物敏感细胞和土壤农杆菌相互作用 植物受伤产生大量对土壤农杆菌感染敏感的细胞 受伤细胞产生一些低分子量的酚类物质可明显刺激土壤农杆菌Ti质粒上毒性区表达 乙酰丁香酮（AS），α-羟基乙酰丁香酮 Ti质粒毒性区基因激活及T-DNA复合物生成 Insert picture Integration of T-DNA into the chromosome of host plant T-DNA整合到植物染色体上 首先在植物靶DNA上出现一个缺刻(nick) 随着DNA解链，宿主细胞的5’—3’外切酶活性使缺刻扩大成裂口(gap) T-DNA侵入裂口，末端与靶DNA单链上的少数核苷酸配对形成异源二倍体； T-DNA悬挂在外侧的末端被切割除去，T-DNA与靶DNA末端相连； 靶DNA上链出现缺刻，以整合后的T-DNA下链为模板合成T-DNA的第二条链，完成整合过程。 References via NCBI The in