Arabis Pumila ApCBF Gene Being Cloned And (Arabidopsis Thaliana CBF Gene)being Transferred In Tobacco

The nature plants can suffer frequently does, is early salt alkaloid and the low temperature and so on the adverse circumstance forces the harm, causes the plant to lack the water to receive the injury,The growth comes under the suppression, the production brings with the serious quality and the output drop for the crops. These non-biology forces the danger which creates to cause self infliction after becomes the agriculture to increase production,The ecological environment improvement, to realize the sustainable development important limiting factor. Therefore, expounded the plant forces the signal to the environment the reply mechanism, enhances the plant using the genetic engineering method the resistance, has become the future agricultural development the key and the foundational work. CBF transcriptional factor passes with CRT/DRE (C-repeat/ Dehydration responsive element) syn-form functional element union activation low temperature and dehydrated response gene expression,Enhances the plant the resistance. Therefore, the CBF kinds transcriptional factor to be able to synthesize the improvement plant the resistance character, is until now the more ideal resistance cyborg gene.In this study, ApCBF gene was cloned from Arabis pumila of xinjiang by using PCR and chromosome walking. Submitted Genebank and its reported: DQ207404.In this study, what had been used inculding CBF1 and CBF3 of CBF family, its downstream COR15a gene , regulating element CRT/DRE in promoter of COR15a gene, Cloning CBF1 and CBF3 from Arabidopsis thaliana, and Cloning ApCOR15a gene from Arabis pumila of xinjiang. On the basis of plasmid pBI121,being used CaMV35S promoter and SAR iductivity promoter synthesized by PCR, six plant expression vectors had been constructed: pCB111, pCB111-1, pCB112,pCB112-1,pCB113, pCB113-1, and then transformed to Agrobacterium GV3101 by electics. Then plant expression vectors were introduced into tobacco disc.Transformed shoots were selected on solid medium containing Kanamycin.These transgenic plants were identifted by PCR, Southern blot and semi-quantified RT-PCR.The results of PCR indicated that 44-88% transgenic plants were positives. The results of Southern blot showed that most of transformed shoots had the interested gene integrated into the genome of tobacco, semi-quantified RT-PCR indicated that most of transgenic plants have expressed CBF1 gene.This research not only laied a foundation for researching on fouction of CBF1,CBF3,ApCOR15a and SAR iductivity promoter,but also provided a new method for anti-frzeeing, presented theorical significance...