水稻CRE基因的克隆、分析及其高效表达载体的构建

Clone and analysis of cytokinin receptor gene along with highly efficient expression vector construction

基于GenBank中公布的水稻(Oryza sativa)基因组序列信息,设计特异引物,结合RT-PCR技术克隆水稻细胞分裂素受体(CTK response,CRE)基因的cDNA全长序列,并利用生物信息学工具进行初步分析。通过限制性内切酶将目的基因片段连接在载体pCAMBIA 1390-35S上,重组质粒分别通过PCR检测和酶切鉴定,成功获得水稻CRE基因的高效表达载体,可直接用于水稻的遗传转化研究。

The sequence of CTK receptor（CRE） gene was obtained based on genome information of Oryza sativa in GenBank.Specific primers were designed and the full length of CRE gene was cloned with with RT-PCR techniques.Then it was subjected to bioinformationally analysis and digested with two restriction enzyme and reconstructed into vector 1 390 with a 35S promoter from cauliflower.The reconstructed vector was detected with PCR amplification and restriction enzyme digestion which together indicted the expression vector had been successfully built and it could directly be used in Oryza sativa genetic engineering.