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多枝赖草谷胱甘肽还原酶基因的克隆及分析
史仁玖1, 赵茂林2, 杨 清3
1.泰山医学院 生物化学与分子生物学教研室;2.北京农业生物技术研究中心;3.南京农业大学 生命科学学院
摘要:
以盐胁迫处理的多枝赖草(Leymus multicaulis)植株新鲜叶片为材料,根据其他植物谷胱甘肽还原酶(GR)氨基酸保守区序列设计简并引物.通过RT—PCR扩增到1个408bp的cDNA片段(Genbank注册号为AY781786);利用DNAMAN软件将5′和3′ RACE获得的5′和3′端序列,拼接成1个全长1580bp的cDNA序列,其包含1个1140bp的开放阅读框架,该阅读框架编码380个氨基酸;多枝赖草谷胱甘肽还原酶氨基酸序列与其他植物的同源性为77%~92%,定量PCR结果表明,GR基因的表达随着盐胁迫时间和盐浓度的增加而加强。
关键词:  多枝赖草  谷胱甘肽还原酶  盐胁迫  GR基因的表达
DOI:
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基金项目:国家自然科学基金项目(30370905);北京市自然科学基金项目(5032009)
Cloning and analysis of glutathione reductase gene from Leymus multicaulis
Abstract:
According to amino acid sequence from glutathione reductase in other plants,a pair of degenerate primers were designed and a 408 bp(Genebank Accession number:AY781786)cDNA fragment was obtained by RT-PCR from Leymus multicaulis under salt stress,5'cDNA end and 3'cDNA end was obtained by RACE.The total cDNA of gene was 1 580 bp in length,including an open reading,encoding 380 amino acids.Its sequences shared high similarity at polypeptide level with the sequences from the plants of Oryza saliva,Zea mays,Vitis vinifera,Zinnia elegans,Nicotiana tabacum,Soybean and Glycine max.RT-PCR analysis indicated that the gene was up-regulated under salt stresses.
Key words:  Leymus multicaulisglutathione reductase  salt stress  GR gene expression

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