水稻耐盐相关基因ZRP4的克隆及其植物表达载体的构建

Cloning salt-tolerance related gene ZRP4 of rice and construction of its plant expression vector

采用RT-PCR技术克隆了水稻O-甲基转移酶ZRP 4基因的cDNA片段,并进行了序列测定与分析。结果表明,该序列全长1 101 bp,编码366个氨基酸,与G enB ank中水稻ZRP 4基因序列比对,核苷酸同源性为99.8%,氨基酸同源性为99.7%。将克隆片段插入中间载体pBPFΩ7,经H indⅢ酶切回收带有P 35s启动子和nos终止子片段,连接pCAM B IA 1301载体,成功构建ZRP 4基因的植物表达载体。

Utilizing RT-PCR,the cDNA of fragment of O-methlytransferanse ZRP4 gene in rice was successfully amplified,which was about 1 101 bp in length and encoded 366 amino acids,and was cloned and sequenced. DNA sequence analysis indicated that the cloned fragment shared high homogeneity to the cDNA of ZRP4 gene in rice. The cloned cDNA of ZRP4 gene was introduced into a middle vector pBPFΩ7. The fragment with P35s promoter and nos were digested by Hind Ⅲ , which were ligated into pCAMBI- A1301 vector,giving an plant expression vector containing ZRP4 gene.