【摘要】 目的:对拟南芥CBF4基因序列进行克隆。方法:用限制性内切酶将CBF4基因从pMD18-T CBF4载体上切下,定向连接到含超强启动子的pC2301-35S-OCS表达载体上,成功构建了CBF4基因植物表达载体pC2301-35S-OCS-CBF4。利用冻融法将此表达载体导入只含辅助质粒的根癌农杆菌中,提取转化质粒,经PCR扩增和酶切验证鉴定表明。结果:CBF4基因植物表达双元载体构建成功。结论:转CBF4基因烟草的抗寒性比野生型烟草要高。更多还原

【Abstract】 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.更多还原