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在本实验室前期研究的干旱胁迫相关的抑制差减文库(SSH)中,发现一个玉米干旱胁迫诱导表达的EST序列,与拟南芥AtCKS2和AtCKS1基因有较高的同源性。应用生物信息学手段和同源克隆的方法,分离得到该基因,命名为ZmCKS2。序列分析表明该基因开放阅读框区267bp,编码88个氨基酸。ZmCKS2蛋白含有真核生物中高度保守的CKS(cyclin-dependent kinase regulatory subunit)结构域。应用在线分析软件预测该基因上游2kb序列表明,该序列具有启动子的核心序列和增强子序列,同时还具有与干旱等多种逆境胁迫相关的调控序列。应用实时荧光定量PCR分析表明该基因在幼穗中表达量最高,且受干旱和MeJA诱导上调表达,受低温和外源ABA诱导下调表达。
In our previous study, we found a EST sequence induced by drought stress in Arabidopsis thaliana and had high homology with AtCKS2 and AtCKS1 genes in Arabidopsis thaliana. Using bioinformatics methods and homologous cloning method, the gene was isolated and named ZmCKS2. Sequence analysis showed that the open reading frame of this gene was 267 bp, encoding 88 amino acids. The ZmCKS2 protein contains a highly conserved CKS (cyclin-dependent kinase regulatory subunit) domain in eukaryotes. Prediction of the 2kb sequence upstream of the gene using on-line analysis software indicated that the sequence has promoter and enhancer sequences, and also has regulatory sequences related to drought stress and other stress conditions. The results of real-time PCR showed that the gene was expressed highest in panicles and induced by drought and MeJA, down-regulated by low temperature and exogenous ABA.