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2025年4月5日 星期六
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甘蔗Lea基因cDNA全长克隆及表达特性
阙友雄 1,刘金仙2,吴嘉云2,许莉萍2,陈如凯2
1. 福建农林大学 农业部甘蔗生理生理生态与遗传改良重点实验室
2. 福建农林大学 农业部甘蔗遗传改良重点开放实验室
Molecular cloning of sugarcane Lea gene and its expression character
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摘要 本研究从甘蔗叶片全长cDNA文库中获得了甘蔗Lea基因cDNA全长序列,命名为Sc-Lea。该基因全长921 bp, 含有一个495 bp编码164个氨基酸的的开放阅读框(open reading frame, ORF), 5’端非编码区(untranslated region,UTR)长64 bp,3’端UTR长362 bp,且在3’端UTR有明显的polyA结构, 起始密码子周围为CCCCAGCCATGGC, -3、+4位为G,C是该段序列的偏好碱基,符合Kozak规则。目的基因原核表达产物分子量大小约为17.9kD。同时还利用定量PCR技术研究了该基因在水杨酸(SA)、H2O2 PEG和NaCl等四种外源非生物胁迫因子作用下的表达特性。研究结果表明,甘蔗Lea基因的表达,会受到PEG和NaCl的强烈诱导,以及H2O2的抑制,同时也会受SA的影响。推测甘蔗Lea基因在甘蔗的抗旱和抗盐机制中发挥作用。
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阙友雄
刘金仙
吴嘉云
许莉萍
陈如凯
关键词 甘蔗LEA蛋白原核表达定量PCR    
Abstract:Late embryogenesis abundant (LEA) protein gene is an important gene in plant stress response. In this paper, a full-length cDNA sequence of sugarcane Lea gene was obtained from sugarcane leaf full-length cDNA library through sequencing and validated by the corresponding bioinformatics analysis , termed Sc-Lea . The full-length of Sc-Lea is 921 bp, with a 495 bp open reading frame(ORF), with a 64 bp 5’ untranslated region(UTR) and 362 bp 3’ UTR . At the 3’ end UTR, it has a clear polyA structure, the sequence around the start codon is CCCCAGCCATGGC,and the base of -3、+4 is G and C respectively which is line with the rules of Kozak. Then, the prokaryotic expression vector, which contained the ORF of sugarcane Lea gene, was constructed and the targeted protein at the molecular weight 17.9kD coding by sugarcane Lea gene was induced by IPTG. Moreover, in order to explore the expression characters of sugarcane Lea gene under the stress of SA, H2O2, PEG and NaCl, the Real-time qPCR approach was applied. The results showed that the expression of the sugarcane Lea gene was induced both by PEG and NaCl, while inhibited by H2O2. Also, the expression of this gene could be influenced by SA. It could be to some extent inferred that the sugarcane Lea gene obtained in this study plays an important role in the sugarcane drought-tolerant and salt-tolerant mechanism.
收稿日期: 2008-11-20     
通讯作者: 阙友雄    
引用本文:   
阙友雄 1,刘金仙2,吴嘉云2,许莉萍2,陈如凯2. 甘蔗Lea基因cDNA全长克隆及表达特性[J]. , 2009, 17(5): 836-842.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2009/V17/I5/836
 
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