Construction of CBF4 Gene in Plant Expression Binary Vector

Objective:CBF4 gene was cloned and then was cut from pMD18-T-CBF4 vector with restraction enzymes.Method:The fragment of CBF4 gene in pC2301-35s-OCS was directively ligated,The plant expression vector pC2301-35s-OCS CBF4 was constructed.The plant expression vector was transformed into Agrobacterium tumefaciens which only contained assistant plasmid in order to construct double-plasmid expression vector.The binary expression vector plasmid was identified by PCR reaction and restriction endonuclease digestion.Result:showed that the plant expression binary vector was constructed successfully.Conclusion:The cold tolerance of transgenic tobacco level was enhanced.