白颖苔草热激转录因子基因的克隆及其序列分析

doi:10.11733/j.issn.1007-0435.2011.06.024

›› 2011, Vol. 19 ›› Issue (6): 1030-1035.DOI: 10.11733/j.issn.1007-0435.2011.06.024

白颖苔草热激转录因子基因的克隆及其序列分析

孙彦, 党卫玲, 杨春华, 邸超, 周禾

中国农业大学动物科技学院草业科学系草业科学北京市重点实验室, 北京100193

收稿日期:2011-06-10 修回日期:2011-09-13 出版日期:2011-12-15 发布日期:2012-07-09
通讯作者: 周禾,E-mail:Zhouhe@cau.edu.cn
作者简介:孙彦(1965),女,内蒙古通辽人,副教授,博士研究生,主要从事草坪科学与管理及草种子检验方面的研究,E-mail:ctsoffice@yahoo.com.cn
基金资助:
草业科学北京市重点实验室开放课题项目资助

Cloning and Sequence Analysis of A Heat Shock Transcription Factor Gene from Carex rigescens

SUN Yan, DANG Wei-ling, YANG Chun-hua, DI Chao, ZHOU He

Department of Grassland Science, College of Animal Science and Technology, China Agricultural University, Key Laboratory of Grassland Science in Beijing, Beijing 100193, China

Received:2011-06-10 Revised:2011-09-13 Online:2011-12-15 Published:2012-07-09

摘要/Abstract

摘要： 白颖苔草(Carex rigescens(Franch.)V.Krecz.)的热激转录因子基因是重要的耐热候选基因。通过序列多重比对设计简并引物,扩增出白颖苔草的热击转录因子基因cDNA的中间片断,并在此基础上利用RACE技术得到3′端cDNA片段和5′端cDNA片段。结果表明:通过序列拼接获得该基因的全长cDNA(1226bp),命名为CrHSF。序列分析表明其含有一个完整的开放阅读框(103～1023bp),编码蛋白质含有306个氨基酸,分子量为38.75kDa,等电点为5.85。CrHSF的氨基酸序列与OsHSF的同源性为59%,与已知的其他热激转录因子也具有较高的同源性。在CrHSF的氨基酸序列中没有预测到信号肽以及跨膜区域,但是具有多种翻译后修饰的加工位点。

关键词: 白颖苔草, 热激转录因子, RT-PCR, RACE, 基因克隆, 序列分析

Abstract: The HSF gene cloned from Rigesecent Sedge(Carex rigescens(Franch.)V.Krecz.) is an important heat-resistant candidate gene.A pair of degenerate primers designed against the conserved regions was used to amplify a cDNA segment(189 bp) from the Rigescent Sedge by RT-PCR.Based on the cDNA segment sequence,primers of 3′ RACE were designed and a 902 bp cDNA segment was acquired.The primers of 5′ RACE were designed from 3′ end sequence of the segment and a 323 bp cDNA segment was acquired.The full-length cDNA(identified CrHSF,1226 bp) was gained by overlapping sequences.The sequence analysis indicates that it contains an open-reading frame from 103 bp to 1023 bp,encoding 306 amino acid residues,with 34.93 kDa of predicted molecular mass and 5.77 isoelectric points.The deduced amino acid sequence shows 59% similarity to OsHSF and it also shares significant similarities with other characterized heat shock transcription factor.In addition,the CrHSF contains no predicted signal peptide,but it contains several post-translation modification and processing sites.

Key words: Carex rigescens(Franch.)V.Krecz., Heat shock transcription factor, RT-PCR, RACE, Molecular cloning, Sequence analysis

中图分类号:

Q943.2

孙彦, 党卫玲, 杨春华, 邸超, 周禾. 白颖苔草热激转录因子基因的克隆及其序列分析[J]. , 2011, 19(6): 1030-1035.

SUN Yan, DANG Wei-ling, YANG Chun-hua, DI Chao, ZHOU He. Cloning and Sequence Analysis of A Heat Shock Transcription Factor Gene from Carex rigescens[J]. , 2011, 19(6): 1030-1035.

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白颖苔草热激转录因子基因的克隆及其序列分析

Cloning and Sequence Analysis of A Heat Shock Transcription Factor Gene from Carex rigescens

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