3种芸薹属植物ROH1基因的克隆及分析

Cloning and Analysis of ROH1 from Three Brassica Plants

[目的]通过PCR和RT-PCR技术从甘蓝、白菜和萝卜中扩增ROH1基因序列,分析该基因的基本特性。[方法]采用生物信息学的相关方法进行分析。[结果]甘蓝、白菜、萝卜ROH1基因没有内含子,甘蓝ROH1基因编码398个氨基酸,白菜和萝卜ROH1基因均编码400个氨基酸,3种植物的ROH1蛋白序列与拟南芥已经公布的ROH1(At1g63930)蛋白氨基酸缺少一段连续的氨基酸序列;ROH1属于DUF793蛋白超家族,它没有跨膜结构域和信号肽,二级结构主要由α-螺旋和无规则卷曲构成。同源比对及功能联想分析发现植物中ROH1非常保守,系统发育分析表明甘蓝、白菜和萝卜的关系最近;结构功能分析发现ROH1可能与BYPASS蛋白和AT4G11300蛋白的功能相似。[结论]为研究DUF793蛋白超家族功能提供参考。

[Objective]The sequences of ROH1 genes were amplified by PCR and RT-PCR from three Brassica plants which are Brassica oleracea,Brassica rape and Raphanus sativus. [Method]Using relevant methods of bioinformatics analyzes. [Result] The results indicated that the g DNA of ROH1 was without introns,ROH1 gene encodes 398 amino acids in Brassica oleracea when it encodes 400 amino acids in the Brassica rape and Raphanus sativus. A continuous period of amino acids was deleted from the ROH1 in the three plants which were compared to ROH1（ At1g63930） in Arabidopsis thaliana; ROH1 belonged to DUF793 protein superfamily,and it might not be a transmembrane domain with no a signal peptide. The secondary structure of ROH1 mainly composed of α-helix and random coil constituted. Homologous alignment and function of the association analysis showed that ROH1 was very conservative,and phylogenetic analysis showed that there were closely relationship between Brassica oleracea,Brassica rape and Raphanus sativus; The structure and function analysis found that ROH1 may resemble to BYPASS protein and AT4G11300 protein. [Conclusion] The study provided a reference for the study of function about DUF793 protein super family.