Cloning of CBF3 Gene from Arabidopsis thaliana and Construction of Plant Expression Vector

[Objective] The aim was to clone CBF3 gene from Arabidopsis thaliana and construct plant expression vector pCAMBIA1301-Rd29A-CBF3.[Method] CBF3 gene and stress-inducible promoter Rd29A were amplified from the genomic DNA of A.thaliana for the construction of plant expression vector.[Result] Sequencing results showed that the cloned CBF3 gene had 750 bp,and showed 100% identity with the sequence published on GenBank.The promoter Rd29A had 1 425 bp,and showed 100% identity with the sequence published on GenBank.[Conclusion] Based on the binary vector pCAMBIA1301,the plant expression vector pCAMBIA1301-Rd29A-CBF3 was constructed successfully,which could materially improve the salt resistance,drought-tolerance,cold resistance of plants.

拟南芥冷诱导型启动子CBF3的克隆及活性检测

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。