Cloning and Characterization of a GDP Dissociation Inhibitor Protein Gene from Medicago sativa L. and Expression in Tobacco

doi:10.11733/j.issn.1007-0435.2011.01.027

›› 2011, Vol. 19 ›› Issue (1): 157-163.DOI: 10.11733/j.issn.1007-0435.2011.01.027

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Cloning and Characterization of a GDP Dissociation Inhibitor Protein Gene from Medicago sativa L. and Expression in Tobacco

WU Yan-hong^1,2, MI Fu-gui¹, LI Zhi-ming², LUAN Shou-quan², CHEN Ling-ling², NR Ri-su², LIANG Qing-wei²

1. Inner Mongolia Agricultural University, Huhhot, Inner Mongolia Autonomous Region 010019, China;
2. Chifeng Academy of Agricultural and Animal Husbandry, Chifen, Inner Mongolia Autonomous Region 024031, China

Received:2010-11-10 Revised:2010-12-07 Online:2011-02-15 Published:2011-02-15

紫花苜蓿GDP解离抑制蛋白基因cDNA的克隆、分析及烟草转化

乌艳红^1,2, 米福贵¹, 李志明², 栾守泉², 陈玲玲², 娜日苏², 粱庆伟²

1. 内蒙古农业大学, 呼和浩特, 010019;
2. 赤峰市农牧科学研究院, 赤峰, 024031

通讯作者: 米福贵,E-mail:mfgui@yahoo.com.cn
作者简介:乌艳红(1967- ),女,蒙古族,内蒙古赤峰市人,博士研究生,研究员,主要从事牧草及帚用高粱种质资源研究与新品种选育研究,E-mail:xuyanwuyanhong@sina.com
基金资助:
现代农业产业技术体系建设专项资金资助

Abstract

Abstract: A cDNA core region of alfalfa GDP dissociation inhibitor protein Gene named MsGDI1 was isolated by RT-PCR.Sequence analysis reveals that the full length of the core region is 1575 bp and contains 1332 bp open reading frame(444 amino acids).Semi-quantitative RT-PCR analysis show that the expression level of MsGDI1 increases significantly with salt-or Aluminum-stress increasing.This result indicates that MsGDI1 may play an important role in both salt and Aluminum tolerances of Medicago sativa L..The MsGDI1 gene is introduced into tobacco by Agrobacterium mediated transformation system using overexpression vector.PCR analyses show that transgenic tobaccos contain the MsGDI1 gene.The function of MsGDI1 gene will be further studied.

Key words: Medicago sativa L., GDP dissociation inhibitor protein gene, RT-PCR, Expression analysis, Overexpression vector construction, Tobacco transformation

摘要： 采用RT-PCR方法,克隆得到紫花苜蓿(Medicago sativa L.)GDP解离抑制蛋白基因cDNA核心区域序列,命名为MsGDI1。序列分析结果表明,该cDNA核心区域全长1575 bp,包含一个1332 bp的最大开放阅读框,编码444个氨基酸。半定量RT-PCR分析结果表明,在盐胁迫和铝胁迫条件下,MsGDI1基因的表达随着胁迫时间的增加而上升,该基因可能与紫花苜蓿抗盐耐铝机理有关。借助于新构建的该基因超表达载体,用农杆菌介导方法转化烟草(Nicotiana tabacum L.)。转基因烟草植株PCR检测的结果表明,MsGDI1基因已经成功插入到烟草基因组中,为进一步研究该基因的功能奠定了基础。

关键词: 紫花苜蓿, MsGDI1(GDP解离抑制蛋白基因), RT-PCR, 表达分析, 超表达载体构建, 烟草转化

CLC Number:

Q756

WU Yan-hong, MI Fu-gui, LI Zhi-ming, LUAN Shou-quan, CHEN Ling-ling, NR Ri-su, LIANG Qing-wei. Cloning and Characterization of a GDP Dissociation Inhibitor Protein Gene from Medicago sativa L. and Expression in Tobacco[J]. , 2011, 19(1): 157-163.

乌艳红, 米福贵, 李志明, 栾守泉, 陈玲玲, 娜日苏, 粱庆伟. 紫花苜蓿GDP解离抑制蛋白基因cDNA的克隆、分析及烟草转化[J]. , 2011, 19(1): 157-163.

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Cloning and Characterization of a GDP Dissociation Inhibitor Protein Gene from Medicago sativa L. and Expression in Tobacco

紫花苜蓿GDP解离抑制蛋白基因cDNA的克隆、分析及烟草转化

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