水稻去泛素化酶OsUCH-L5基因的克隆及表达分析

Cloning and Expression Analysis of OsUCH-L5 Gene Encoded Rice Deubiquitin Enzyme

通过生物信息学及RT-qPCR(quantitative Real-time PCR,RT-qPCR)的方法对水稻OsUCH-L5基因进行探究。生物信息学预测结果表明,该基因全长837 bp,编码278个氨基酸;OsUCH-L 5蛋白的相对分子质量为31.64 kDa,等电点为5.66,为亲水性蛋白,具备Peptidase_C 12及UCH_C结构域;系统进化树表明,与二穗短柄草(Brachypodium distachyon)BdUCH-L 5亲缘关系最近。RT-qPCR结果显示,OsUCH-L5基因具有组织特异性,在根中表达最佳;在不同干旱脱落酸(200μmol·L^(-1))、D-甘露醇(1 mmol·L^(-1))及氯化钠盐胁迫(250 mmol·L^(-1))处理下,基因表达量存在显著差异,并分别在0.5 h、6 h、12 h时表达水平最佳;在油菜素内酯(200μmol·L^(-1))处理下,在3 h时OsUCH-L5基因表达量达到最大值;同时,OsUCH-L5基因还受4℃低温和40℃高温的诱导表达,即分别在3 h和1 h时基因表达量达到最大值。因此该研究为进一步探索水稻UCH家族在调节水稻抗性、信号转导、营养以及盐分胁迫调控等过程中的作用提供基础。

In this study,the rice OsUCH-L5 gene was investigated based on bioinformatics and quantitative Real-time PCR(RT-qPCR).The results of bioinformatics showed that the total length of the gene was 837 bp,encoding 278 amino acids.The relative molecular weight of OsUCH-L 5 protein was 31.64 kDa,and the isoelectric point was 5.66.OsUCH-L 5 protein belonged to hydrophilic protein and possessed Peptidase_C 12 and UCH_C domains.The phylogenetic tree indicated that OsUCH-L 5 protein was the most closely related to Brachypodium distachyon UCH-L 5.RT-qPCR results showed that OsUCH-L5 gene was tissue-specific,especially significant expressed in roots.Under different drought treatments with abscisic acid(200μmol·L^(-1)),D-mannitol(1 mmol·L^(-1))and sodium chloride(250 mmol·L^(-1)),the gene expression levels were significantly different,and the maximum expression levels were found at 0.5 h,6 h and 12 h,respectively.Under the treatment with 200μmol·L^(-1)brassinolide,the expression of OsUCH-L5 gene reached the maximum at 3 h,and the expression of OsUCH-L5 gene was also induced by the low temperature of 4℃and high temperature of 40℃,that was,the gene expression reached the maximum at 3 h and 1 h respectively.Therefore,this study provided a basis for further exploration of the role of UCH family in regulating rice resistance,signal transduction,nutrition and salt stress regulation.