东方山羊豆水通道蛋白基因的克隆及初步分析

doi:10.11733/j.issn.1007-0435.2011.02.026

›› 2011, Vol. 19 ›› Issue (2): 331-339.DOI: 10.11733/j.issn.1007-0435.2011.02.026

东方山羊豆水通道蛋白基因的克隆及初步分析

吴晓静^1,2, 王学敏², 高洪文², Dzyubenko Nikolay³, Chapurin Vladimir³, 刘大林¹

1. 扬州大学动物科学与技术学院, 扬州, 225009;
2. 中国农业科学院北京畜牧兽医研究所, 北京, 100193;
3. 俄罗斯瓦维洛夫全俄植物栽培研究所, 圣彼得堡, 190000

收稿日期:2010-11-15 修回日期:2011-03-01 出版日期:2011-04-15 发布日期:2011-04-15
通讯作者: 高洪文,E-mail:gaohongwen@263.net
作者简介:吴晓静,(1985- ),女,内蒙古包头人,硕士研究生,主要研究方向为牧草种质资源与育种,E-mail:xiaojing_wu09@yahoo.com.cn
基金资助:
现代农业产业技术体系建设专项资金;中国农业科学院基本科研业务费(2011cj-16);十二五科技支撑(20011BAD17B01)资助

Cloning and Analysis of the Aquaporins Gene from Galega Orientalis

WU Xiao-jing^1,2, WANG Xue-min², GAO Hong-wen², Dzyubenko Nikolay³, Chapurin Vladimir³, LIU Da-lin¹

1. College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu Province 225009, China;
2. Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China;
3. N.I.Vavilov All-Russian Research Institute of Plant Industry, St.Petersburg 190000, Russia

Received:2010-11-15 Revised:2011-03-01 Online:2011-04-15 Published:2011-04-15

摘要/Abstract

摘要： 根据已知EST片段结合RT-PCR和RACE技术,从东方山羊豆(Galega Orientalis)中克隆到一个水通道蛋白(AQP)基因,命名为GoAQP(GenBank登录号:HM 803185)。测序和生物信息学分析表明,此序列全长1258bp,开放读码框(ORF)870 bp,编码289个氨基酸。GoAQP包含MIP家族信号序列HINPAVT/SFG,高等植物PIP高度保守序列GGANXXXXGY和TGI/TNPARSL/FGAAI/VI/VF/YN。与拟南芥AQP基因家族的系统进化分析表明,GoAQP属于PIP1亚型AQP蛋白。Real-Time PCR检测结果表明,GoAQP基因在根中表达量最高,茎中表达量最少;且受NaCl和PEG胁迫表达上调,NaCl表达呈双峰分布。因此GoAQP基因可能参与了东方山羊豆耐盐性调控。同时成功构建超表达载体pCAMBIA-1302-GoAQP,为转基因验证基因功能创造了基础。

关键词: 东方山羊豆, 水通道蛋白基因, 表达分析, 表达载体构建

Abstract: Based on a deposited EST fragment,the full-length cDNA of one novel gene encoding aquaporin had been cloned from Galega Orientalis using RT-PCR and RACE technology.The new gene was identified as the GoAQP gene and had been assigned GenBank accession number: HM803185.Bioinformatics analysis showed that the full-length cDNA of this gene is 1258 bp including an 870-bp open reading frame(ORF),which encodes a protein of 289 amino acids.GoAQP protein possesses the conserved MIP family signal sequence HINPAVT/SFG,the highly conserved sequence of the higher plant GGANXXXXGY and TGI/TNPARSL/FGAAI/VI/VF/YN.Compared to the AQP family from Arabidopsis thaliana,phylogenetic analysis indicated that GoAQP belong to PIP1 subfamily.Real-Time PCR results revealed that expression of GoAQP was the most abundant in root and the least in stem.The expression level of GoAQP in leaves were increased with NaCl and PEG stress,and showed a bimodal distribution pattern in NaCl treatment.This suggested that GoAQP might be involved in the manipulation of salt tolerance of Galega Orientalis.The expression plasmid pCAMBIA-1302-GoAQP was constructed for the further gene function analysis using the transgenic technology.

Key words: Galega Orientalis, Aquaporin, Expression analysis, Recombinant expression plasmid

中图分类号:

Q785

吴晓静, 王学敏, 高洪文, Dzyubenko Nikolay, Chapurin Vladimir, 刘大林. 东方山羊豆水通道蛋白基因的克隆及初步分析[J]. , 2011, 19(2): 331-339.

WU Xiao-jing, WANG Xue-min, GAO Hong-wen, Dzyubenko Nikolay, Chapurin Vladimir, LIU Da-lin. Cloning and Analysis of the Aquaporins Gene from Galega Orientalis[J]. , 2011, 19(2): 331-339.

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东方山羊豆水通道蛋白基因的克隆及初步分析

Cloning and Analysis of the Aquaporins Gene from Galega Orientalis

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