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。
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.
出处
《生物技术》
CAS
CSCD
北大核心
2009年第4期6-9,共4页
Biotechnology
基金
国家烟草专卖局科技计划项目(No.110200601010)资助
