拟南芥冷诱导型启动子CBF3的克隆及活性检测

Cloning of Cold-Induced Promoter CBF 3 from Arabidopsis thaliana and Its Activity Detection in Transgenic Tobacco

目的:构建冷诱导型启动子CBF3基因的植物表达载体,并将其转入烟草。方法:以拟南芥基因组DNA为模板,通过特异PCR扩增,克隆冷诱导表达启动子CBF3(C-repeat binding factor)。用CBF3启动子替换pBI121载体上的35S启动子构建新的载体pBC-GUS,通过农杆菌介导的叶盘法转化烟草。结果:获得了转基因烟草,转基因烟草的GUS组织化学染色及PCR分析结果表明,在低温诱导下,CBF3启动子可增强GUS基因表达。结论:CBF3启动子可应用于植物抗冷基因工程研究。

Objective：The expression vector of pBC-GUS was constructed and transferred into tobacco to get the transgenic plants.Method：Cold-induced promoter CBF 3（C-repeat binding factor）was amplified and cloned by PCR from the genomic DNA of the Arabidopsis thaliana.Promoter 35S of vector pBI121 was replaced by promoter CBF 3,35S of vector pBI121 was replaced by promoter CBF 3,thus vector pBC-GUS was constructed and transferred into tobacco by co-culturing excised cotyledon explants with Agrobacterium tumefaciens,and transgenic plants were obtained.Result：PCR and GUS histochemical stain of transgenic tobacco showed that promoter CBF 3 induced by low temperature stress strengthened expression of gene GUS.Conclusion：Therefore,promoter CBF 3 could be used to plant gene engineering for cold resistance improvement.