水稻中一个与COI1同源的基因克隆及其表达分析

Cloning and Expression Analysis of a COI 1 Gene Homologue from Rice

利用同源序列分析 ,在水稻基因数据库中检索到了几条与拟南芥中的COI1基因具有高度同源性的EST(expressionsequencetag)片段和相应的基因组序列 .根据EST拼接和基因组比较结果设计引物 ,利用RT PCR在水稻中得到与COI1高度同源的cDNA ,并在基因组中推断出相应的基因 ,命名为RCOI1.RCOI1与COI1在氨基酸水平有 74 %的同源性 .与COI1相似 ,RCOI1蛋白也具有典型的F box和LRRs结构域 .半定量RT PCR显示该基因的表达受茉莉酸甲酯 (MeJA)的诱导 ,说明这个基因可能参与水稻茉莉酸应答反应 .该基因的发现对于探索单子叶植物的茉莉酸信号转导途径 ,以及研究该途径在农作物中的抗虫抗病和抵抗恶劣环境等方面的作用都有重要意义 .

Using blast analysis,several EST fragments shown high homologue to Arabidopsis gene COI 1,were found in rice gene database,together with a genomic fragment in accordance with these EST fragments. A new gene’s cDNA was isolated from rice,designated as RCOI 1,via EST sequence alignment method,genomic analysis and RT-PCR. The genomic gene of RCOI 1 was obtained. There is 74% homology at amino acid level between RCOI1 and COI1.The protein encoded by RCOI 1 contains characteristic F-box motif and LRRs motif,indicating that it belongs to a kind of special F-box protein.Semi-quantitative RT-PCR demonstrated that the expression of RCOI 1 was clearly regulated by the induction of Methyl Jasmonate.Therefore,the new gene has the putative characteristics of COI 1,and is probably involved in the JA responses of rice.The discovery of RCOI 1 not only provides insight in the study of the jasmonate signal transduction pathway in monocotyledon,but is also helpful in regulating crop’s defense against insects and pathogens,and improving their tolerance to stresses.