引用本文
  • 许洋,雷晨,佘露露,等.沙冬青脱水素基因的分子克隆与序列分析[J].植物遗传资源学报,2019,20(5):1317-1324.    [点击复制]
  • XU Yang,LEI Chen,SHE Lu-lu,et al.Molecular Cloning and Sequence Analysis of The Dehydrin Genes in Ammopiptanthus mongolicus[J].植物遗传资源学报,2019,20(5):1317-1324.   [点击复制]
【打印本页】 【在线阅读全文】【下载PDF全文】 查看/发表评论下载PDF阅读器关闭

←前一篇|后一篇→

过刊浏览    高级检索

本文已被:浏览 1005次   下载 1221 本文二维码信息
码上扫一扫!
沙冬青脱水素基因的分子克隆与序列分析
许洋1, 雷晨1, 佘露露1, 王瑾瑜2, 杨佳慧1, 陆雅宁1, 卢存福1, 陈玉珍1
0
(1北京林业大学生物科学与技术学院/林木育种国家工程实验室/教育部林木花卉遗传育种重点实验室,北京 100083;2清华大学分析测试中心,北京 100084)
摘要:
脱水素(Dehydrin)是晚期胚胎发生蛋白(LEA)家族中最具特色的一组蛋白,对植物抵抗非生物逆境胁迫起到非 常重要作用。本文从抗逆性较强的常绿阔叶灌木沙冬青中克隆出 4 个含有完整 ORF 的脱水素基因:命名为 AmDHN3.2F、 AmDHN5.1F、AmDHN6.2F 和 AmDHN7.2,并进行了相关生物信息学分析。使用 DNAMAN 软件预测结果显示,4 个基因都含有一个 K 片段和 S 片段。理化性质分析表明,4 种脱水素蛋白均为亲水性蛋白,其中 AMDHN3.2F 为碱性蛋白,AMDHN5.1F、AMDHN6.2F 和 AMDHN7.2 为酸性蛋白;理论相对分子质量最大的是 AMDHN7.2(21.42kD),最小的是 AMDHN3.2F(10.69kD);理论等电点最 大的是 AMDHN3.2F(9.01),最小的是 AMDHN6.2F(6.21)。蛋白二级和三级结构预测结果发现,四种蛋白均由三种结构组成: α-螺旋(7.50%-32.63%)、不规则卷曲(55.79%-73.00%)和延伸链(11.58%-19.50%)。采用邻接法构建系统进化树,并结合 同源相似性分析发现,4 种脱水素蛋白均与蒺藜苜蓿 MtDHN3 和拟南芥 AtDHN10 亲缘关系较近。利用 XSTREAM 软件分析结果显 示,AMDHN5.1F、AMDHN6.2F 和 AMDHN7.2 三个蛋白序列具有串联重复单元,且主要集中在脱水素的中间部分。通过 DOTTER 软 件生成点阵图发现,在中间区域存在大量的短片段重复,说明脱水素序列的收缩或扩张主要发生在中间部分。这些研究结果 为进一步开展沙冬青 DHN 基因家族的功能分析奠定了基础。
关键词:  脱水素  沙冬青  基因家族克隆  生物信息学  串联重复
DOI:10.13430/j.cnki.jpgr.20190129002
投稿时间:2019-01-29修订日期:2019-03-06
基金项目:中国国家自然科学基金(31270737);北京市自然科学基金(6112016)
Molecular Cloning and Sequence Analysis of The Dehydrin Genes in Ammopiptanthus mongolicus
XU Yang1, LEI Chen1, SHE Lu-lu1, WANG Jin-yu2, YANG Jia-hui1, LU Ya-ning1, LU Cun-fu1, CHEN Yu-zhen1
(1College of Biological Sciences and Technology, National Engineering Laboratory for Tree Breeding, Key Laboratory of Genetics and Breeding in Forest Tree and Ornamental Plants of Education Ministry, Beijing Forestry University, Beijing 100083;2Analysis and Testing Center,Tsinghua University,Beijing 100084)
Abstract:
Dehydrin is the most distinctive group of proteins in the late embryogenesis protein (LEA) family, which plays an important role in plant resistance to abiotic stress. In this paper, we isolated the complete coding sequences of four dehydrin genes, designated AmDHN3.2F, AmDHN5.1F, AmDHN6.2F and AmDHN7.2, in stress-resistant evergreen broad-leaved shrub Ammopiptanthus mongolicus. The bioinformatics analysis using DNAMAN software suggested that all four genes carried one K and one S domain. The analysis of physicochemical properties showed that all four proteins belonged to hydrophilic proteins. AmDHN3.2F was alkaline protein, while AmDHN5.1F, AmDHN6.2F and AmDHN7.2 were acidic proteins. AmDHN7.2 exhibited the largest theoretical relative molecular weight (21.42 KD), and AmDHN3.2F showed the smallest weight (10.69KD). AmDHN3.2F exhibited the highest theoretical isoelectric point (PI = 9.01), and AmDHN6.2F showed the smallest isoelectric point (6.21). By in silico prediction of secondary and tertiary structure, the four dehydrin proteins composed of three types of motifs: alpha helix (7.50%-32.63%), irregular curl (55.79%-73.00%) and elongation chain (11.58%-19.50%). The phylogenetic tree constructed by Neighbor-joining method suggested that the putative proteins of four dehydrin genes were related to MtDHN3 and AtDHN10. By XSTREAM software, it was found that the protein sequences of AmDHN5.1F, AmDHN6.2F and AmDHN7.2 contained tandem repeat units, which were enriched in the middle part of dehydrin. According to the dot maps generated by DOTTER software, a large number of short repeats were found in the middle part, implying the location where the shrinkage or expansion of dehydration sequence mainly occurred. This findings lay a foundation for further functional analysis of DHN gene family in Ammopiptanthus mongolicus.
Key words:  dehydrin  Ammopiptanthus mongolicus  gene family cloning  bioinformatics  tandem repeat

用微信扫一扫

用微信扫一扫