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陆地棉(Gossypium hirsutum L.)中一个RanBP2锌指蛋白基因的分子克隆与表达分析

Molecular cloning and expression analysis of a RanBP2 zinc finger protein gene in upland cotton (Gossypium hirsutum L.).

作者信息

Chang Ping-An, Li Biao, Ni Xiao-Min, Xie Yong-Fang, Cai Ying-Fan

机构信息

Key Laboratory of Molecular Biology, College of Bio-information, Chongqing University of Posts and Telecommunications, Chongqing 400065, PR China.

出版信息

Colloids Surf B Biointerfaces. 2007 Apr 1;55(2):153-8. doi: 10.1016/j.colsurfb.2006.11.042. Epub 2006 Dec 9.

Abstract

Gossypol is an important resistant substance of Gossypium, and its storage organ is pigment gland. Although, the relationship between gossypol and pigment gland has been studied for a long time, the development mechanism of pigment gland has not been revealed up to now in molecular perspective. On the basis of differentially expressed cDNAs fragments at the stage of the cotton gland development using suppression subtractive hybridization (SSH), the complete cDNA sequence of a novel RanBP2 zinc finger protein (ZFP) gene was cloned by rapid amplification of cDNA ends (RACE) from upland cotton (Gossypium hirsutum L.), Xiangmian 18. The cotton RanBP2 ZFP cDNA (GenBank accession number: DQ173926) is 717 base pair (bp) long with an open reading frame encoding 139 amino acids, which encodes a 15.6 kDa protein. The cotton RanBP2 ZFP had three Ran-binding protein (RanBP) two zinc finger motifs and belonged to RanBP2 ZFP family. There is a 292-base non-coding sequence at 3' cDNA end, which includes polyA sequence. Sequence alignment analysis revealed that the cDNA nucleotide and its deduced amino acid sequence are moderately identical to the putative ZFP from other species. The mRNA expressing profiles of the novel ZFP gene was investigated by reverse transcription-polymerase chain reaction (RT-PCR). The result showed that it expressed at different development stages of gland, including the undeveloped stage, developing stage, developed stage and cotyledon stage. However, with the development of pigment gland, the mRNA levels in the gland-developed seed and cotyledon were increased to about 1.5 and 2 folds of that in gland-undeveloped seed, respectively, which suggested that the novel ZFP played a role in the development of the cotton gland.

摘要

棉酚是棉花的一种重要抗性物质,其储存器官是色素腺体。尽管棉酚与色素腺体之间的关系已被研究了很长时间,但从分子角度来看,色素腺体的发育机制至今尚未揭示。基于抑制性消减杂交(SSH)技术获得的棉花腺体发育阶段差异表达的cDNA片段,通过cDNA末端快速扩增(RACE)技术从陆地棉(Gossypium hirsutum L.)湘棉18中克隆到一个新的RanBP2锌指蛋白(ZFP)基因的全长cDNA序列。该棉花RanBP2 ZFP cDNA(GenBank登录号:DQ173926)长717碱基对(bp),具有一个编码139个氨基酸的开放阅读框,编码一个15.6 kDa的蛋白质。该棉花RanBP2 ZFP具有三个Ran结合蛋白(RanBP)两个锌指基序,属于RanBP2 ZFP家族。在cDNA 3'末端有一个292碱基的非编码序列,其中包括polyA序列。序列比对分析表明,该cDNA核苷酸及其推导的氨基酸序列与其他物种的假定ZFP有中度同源性。通过逆转录-聚合酶链反应(RT-PCR)研究了该新ZFP基因的mRNA表达谱。结果表明,它在腺体的不同发育阶段均有表达,包括未发育阶段、发育阶段、成熟阶段和子叶阶段。然而,随着色素腺体的发育,腺体成熟种子和子叶中的mRNA水平分别增加到腺体未发育种子中的约1.5倍和2倍,这表明该新ZFP在棉花腺体发育中发挥作用。

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