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当前位置: 首页 > 文章 > 紫花苜蓿抗逆基因MsDUF的克隆及其功能分析 中国农业科学 2013,46 (2) 424-432
Position: Home > Articles > Isolation and Functional Analysis of the Stress Resistance Gene MsDUF in Medicago Sativa L. Scientia Agricultura Sinica 2013,46 (2) 424-432

紫花苜蓿抗逆基因MsDUF的克隆及其功能分析

作  者：
韩博;王卫栋;杨培志;张攀;呼天明
单  位：
西北农林科技大学动物科技学院
关键词：
紫花苜蓿;MsDUF;高渗胁迫;激素诱导;亚细胞定位;功能分析
摘  要：
【目的】克隆紫花苜蓿(Medicago sativa L.)抗逆新基因MsDUF,并对其进行序列特征分析,了解该基因在逆境胁迫下的表达模式。【方法】利用RACE法获得紫花苜蓿MsDUF全长的cDNA序列,对该序列进行生物信息学分析;用基因枪法进行MsDUF的亚细胞定位分析;采用实时荧光定量PCR研究该基因在高含量NaCl和PEG-6000的胁迫下,以及ABA和GA3诱导下的表达模式。【结果】测序结果显示,该基因cDNA全长714 bp,包含一个633 bp的完整开放阅读框,编码210个氨基酸,命名为MsDUF,GenBank登录号为JX183734。氨基酸BLASTP分析表明,MsDUF氨基酸序列与拟南芥、大豆和蒺藜苜蓿等具有53%—98%的相似性,属于DUF4228 superfamily。实时荧光定量PCR研究表明,该基因在逆境胁迫下上调表达。【结论】从紫花苜蓿中克隆了抗逆相关基因MsDUF,发现其定位于细胞质中,实时荧光定量PCR结果分析表明该基因在紫花苜蓿的抗逆反应中起着重要作用。
译  名：
Isolation and Functional Analysis of the Stress Resistance Gene MsDUF in Medicago Sativa L.
作  者：
HAN Bo1,2,WANG Wei-dong1,YANG Pei-zhi1,ZHANG Pan1,HU Tian-ming1(1College of Animal Science and Technology,Northwest A&F University,Yangling 712100,Shaanxi;2State Key Laboratory of Crop Stress Biology in Arid Areas,Yangling 712100,Shaanxi)
关键词：
Medicago sativa;MsDUF;hypertonic stress;hormone-induced;subcellular localization;functional analysis
摘  要：
【Objective】This study is aimed to characterize the stress resistance gene MsDUF in Medicago sativa by cloning and analysis of its sequence,and study the function of MSDUF under stress conditions.【Method】The full-length cDNA sequence of MsDUF was isolated by RACE.The obtained cDNA sequence and the deduced amino acid sequence were analysed.Using gene gun in subcellular localization.Real-time quantitative PCR was used to assess the expression of MsDUF in response to NaCl,PEG-6000,ABA and GA3.【Result】The sequencing result showed that the cloned cDNA(designated as MsDUF,GenBank accession No.JX183734) was 714 bp,contains an open reading frame(ORF) of 633bp,and encoding a protein with 210 amino acids.BLASTp search revealed 53% to 98% similarities between the MSDUF protein and those reported MSDUF from other species,such as Arabidopsis thaliana,Glycine max and M.truncatula.The RT-PCR result showed that MsDUF was upregulatedly in response to stress resistance.【Conclusion】The stress resistance related gene MsDUF was cloned from M.sativa and found that it localized in cytoplasm.RT-PCR analysis showed that the gene MsDUF plays an important role in response to the abiotic stress.It is a novel stress resistance gene in M.sativa.

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