Ti质粒与植物基因组的相互作用.pptxVIP

Ti质粒与植物基因组的相互作用会计学What is Ti-plasmid?第1页/共21页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. 第2页/共21页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.第3页/共21页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第4页/共21页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第5页/共21页OCS plasmid (octopine )pTiAch5, pTiA6NC, pTiB653, pTiAg162 NOS plasmid (nopaline )pTiT37, pTiT38第6页/共21页土壤农杆菌-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第7页/共21页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.第8页/共21页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-羟基乙酰丁香酮第9页/共21页Ti质粒毒性区基因激活及T-DNA复合物生成Insert picture第10页/共21页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。第11页/共21页References via NCBIThe in