Molecular cloning of a dehydration-responsive protein gene (MRD22) from mulberry, and determination of abiotic stress patterns of MRD22 gene expression.

Bioorganicheskaia khimiia Pub Date : 2014-03-01
Heng Wang, Zhaoyue Liu, Feng Li, Yuhua Wang, Rongjun Fang, Weiguo Zhao, Long Li
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Abstract

A full-length cDNA sequence coding for Dehydration-responsive protein gene of mulberry tree, which we designated was MRD22 (GenBank accession number: JQ804833) was cloned based on mulberry expressed sequence tags (ESTs). MRD22 is 1503 bp long, contains a 334 bp 5'-UTR (untranslated region) and a 563 bp 3'-UTR, encodes 201 amino acids with a predicted molecular weight of 54.28 kDa and an isoelectric point of 9.35. Phylogenetic analysis based on MRD22 sequences from different species showed that mulberry has close relationship with Populus trichocarpa, Ricinus communis, Camellia sinensis, Gossypium hirsutum, Gossypium barbadense and so on. The expression level of the MRD22 gene under conditions of drought, low temperature and salt stresses was quantified by qRT-PCR. The results show that the expression level changed significantly under the stress conditions compared to the normal growth environment. It helps us to get a better understanding of the molecular basis for signal transduction mechanisms underlying the stress response in mulberry.

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桑树脱水反应蛋白基因MRD22的克隆及非生物胁迫下MRD22基因表达模式的测定。
利用桑树表达序列标签(ESTs)克隆了桑树脱水反应蛋白基因的全长cDNA序列,编号为MRD22 (GenBank登录号:JQ804833)。MRD22全长1503 bp,包含一个334 bp的5'-UTR(非翻译区)和一个563 bp的3'-UTR,编码201个氨基酸,预测分子量为54.28 kDa,等电点为9.35。基于不同种MRD22序列的系统发育分析表明,桑树与毛杨(Populus trichocarpa)、蓖麻(Ricinus communis)、山茶(Camellia sinensis)、绵棉(Gossypium hirsutum)、巴氏绵棉(Gossypium barbadense)等亲缘关系较近。采用qRT-PCR方法定量分析干旱、低温和盐胁迫下MRD22基因的表达水平。结果表明,与正常生长环境相比,胁迫条件下的表达量发生了显著变化。这有助于我们更好地了解桑树胁迫反应的信号转导机制的分子基础。
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