农杆菌介导转化法.pptVIP

缺失研究也表明：右边界重复序列缺失后，T-DNA不能转移，而左边界重复序列的缺失会稍稍降低T-DNA转移频率(Timmerman?B．1988)，这进一步说明T-DNA单链是在从右边界到左边界5’向3‘替换合成中释放出来的。 右边界的缺失，使得DNA合成不能起始，因而T-DNA不能释放，所以T-DNA不能转移到植物细胞。而左边界的缺失，仅会影响DNA合成过程的终止，可能会降低导致T-DNA单链释放，而不会明显影响T-DNA单链的形成。 冠瘿也是在冠瘿碱胞内合并成分泌出来的,被冠瘿碱分解基因分解成根癌农杆菌生长所须的碳源和氮源。 冠瘿碱分解区 Agrobacterium-mediated transformation ? Advantages: 1.???Cleaner inserts 2.???Higher co-expression 3.???More flexible in tissue types(in planta transformation) almost any type of tissues or cells 4. Mostly widely used system ,both monocot and dicot 5. Easy in manipulation , Disadvantages 1.? Only good for nuclear transformation 2.??Limited in host range,genotype-dependent 3.???Detrimental to some tissues Introduction to Agrobacterium Moloecular mechanisms of T-DNA transfer into the plant genome Factors influencing the T-DNA delivery Regeneration and selection of transgenic plants Types A.tumefaciens : the causative agent of plant disease crowng gall. rhizogenes : provokes hair roots. Both crown gall and root hair are neoplastic diseases (plant tumors), both gram-negative soil bacterium. Based on the opines produced in the tumors,Ti plasmids are divided in four groups,while the Ri plasmids fall in three groups. Ti plasmids u???? Octopine: pTiAch5,pTiAg57,or pTi19955 Nopaline: pTiC58,pTi1D135,pTi223 Leucinnopine: pTi542,pTiAT4 D,L-succinamopine: pTiEU6,pTiAT181 Ri plasmids u???? Mannopine: pRi8196,pRiTR7 Agropine: pRi1855,pRi15834,pRiA4 Cucumopine: pRi1659 Strains: Agrobacterium Strains and the origins of Agrobacterium strains(see attached table 1 and 2)refernce plasmid1:238-253(1978) Examples: C58 (C58 chromosome,pTi C58) ABI (C58 chromosome,pTiMp90RK,disarmed TiC58) GV3850 (C58 chromosome,pTiGV3850,disarmed TiC58) A281 (C58 chromosome,pTiBo542) EHA101 (C58 chromosome