张贺翠 廉小平 柳菁 李帮秀 周燕

摘要  [目的]通过PCR和RTPCR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。

关键词 ROH1基因;基因克隆;DUF793蛋白超家族

中图分类号 S188  文献标识码

A  文章编号 0517-6611（2015）03-018-04

Cloning and Analysis of ROH1 from Three Brassica Plants

ZHANG Hecui， LIAN  Xiaoping， LIU Jing et al

（College of Agronomy and Biotechnology ， Southwest University ， Chongqing 400716）

Abstract   [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 gDNA 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 αhelix 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.

Key words  ROH1 gene; Gene clone;  DUF793 protein super family

基金項目 中央高校基本业务费专项经费（XDJK2013C121）

作者简介 张贺翠（1985- ），女，河南滑县人，助理实验师，硕士，从事生物化学与分子生物学方面的研究。

收稿日期 20141125

囊泡分泌和转运过程是真核细胞生命活动的基本事件之一，是神经信号传导和内分泌激素释放的基础，细胞内大多数重要的信号蛋白或者膜蛋白需要借助囊泡的转运和分泌来实现^[1]。囊泡转运和分泌过程中，由多亚基组成的7类复合物被证明在真核细胞的内膜转运和囊泡拴系过程中发挥重要作用。其中研究最为活跃和进展最快的是Exocyst，它是由SEC3、SEC5、SEC6、SEC8、SEC10、SEC15、EXO70 和 EXO84 8亚单位构成^[2]。

Elias等^[3]首先从水稻和拟南芥中发现了EXO70A1，它是EXO70家族中的一员。……