Efficiency of induction of processing of TDNK 48 hours of an incubation of agrobacteria with fabrics of plants. The caption of viable bacteria after an incubation from ekstsudata of plants was checked by their seeding in various cultivations for Petri's cups with the agarizovanny LB environment. Each option of experiences was carried out in three povtornost.
Emergence of this intermediat is found in eight hours after addition of the inductor, and its quantity collects for the next 40 hours more, than twice, and then begins to decline, showing that process is limited.
Night culture of A. tumefaciens of C58C1 (pGV385 which is grown up on a rotational shaker (20 cycles/min.) at 290 C in the liquid LB environment collected centrifugation and suspenzirovat in the environment of MS – 0,1 M the phosphatic buffer ( 5, to acidity of A600 – 0,0
Thus, agrobacteria are natural "gene engineers", the carrying-out transfer of genetic information, amazing on phylogenetic range. On the basis of agrobacteria effective vector systems for genetic engineering of plants are designed.
In a kachetva of positive control used agrobacterial culture with addition as the inductor of 100 microns of an atsetosiringon. As negative control used the agrobacterial cages which are grown up on the Wednesday without atsetosiringon and ekstsudat of plants.
Plants grew up in the greenhouse in vegetative vessels at a temperature of 18-250 With and 12 hour light period. Relative humidity of air in the greenhouse was maintained within 65–70%. For the analysis took sterile fabrics of leaves. Sterilization was carried out sodium hypochloride within 5–10 minutes, and then 3 times washed out material sterile water and used in experiments on induction of vir-genes of Ti-plasmids. For comparison two-submultiple plants of tobacco red and flax of dolgunets were taken.