棉花中一个新型转录因子GhWRI1的表达特征与功能分析

生物技术通报 ›› 2013, Vol. 0 ›› Issue (6): 80-86.

棉花中一个新型转录因子GhWRI1的表达特征与功能分析

李鑫, 王正明, 薛伟, 初明光

(清华大学分子生物学实验室, 北京 100084)

收稿日期:2013-06-20 修回日期:2013-06-20 出版日期:2013-06-20 发布日期:2013-06-20
基金资助:
国家重点基础研究和发展计划(2010CB126003), 国家转基因动物和植物研究项目(2009ZX08005-026B, 2011ZX08009-003, 2011ZX08005-003)

Identification and Characterization of a Novel Gene, GhWRI1, Encoding an AP2-type Transcription Factor in Gossypium hirsutum

Li Xin Wang Zhengming Xue Wei Chu Mingguang

(Laboratory of Molecular Biology and MOE Laboratory of Protein Science, School of Life Sciences, Tsinghua University, Beijing 100084)

Received:2013-06-20 Revised:2013-06-20 Published:2013-06-20 Online:2013-06-20

摘要/Abstract

摘要： AP2(APETALA2) /EREBP(ethylene-responsive element binding protein) 是一类特异性存在于植物中的转录因子, 在植物中的许多代谢过程都起着十分重要的作用。从棉花中分离并获得了一个新的AP2基因GhWRI1, 该基因长1 314 bp, 所编码的蛋白含有437个氨基酸, 分子量约为48 kD, pI为5.77。序列分析结果显示, GhWRI1蛋白含有2个AP2结构域, 该蛋白属于AP2家族;与GFP蛋白的融合表达试验结果显示, 在洋葱表皮细胞中, GhWRI1是特异性在细胞核中表达的。Real-time RT-PCR 数据显示GhWRI1主要在真叶、根、纤维以及种子中表达, 而且其在纤维和种子中的表达量是随着发育时间的推移而不断升高的。同时构建了GhWRI1与LUC共表达的载体, 通过检测LUC的活性来反映GhWRI1的表达量, 试验数据显示, 当GhWRI1与LUC共表达后, LUC的活性提高了约20倍。试验结果都显示GhWRI1是一个很强的转录激活子。

关键词: WRI1基因, 转录因子, AP2结构域, 转录激活试验, 棉花

Abstract: AP2(APETALA2)/EREBP(ethylene-responsive element binding protein)genes comprise a family of plant-specific transcription factors, many of which have been linked to the regulation of essential biological processes. In this study, a novel AP2 gene, named GhWRI1, was isolated from Gossypium hirsutum(cotton). The GhWRI1 cDNA has a total length of 1 314 bp and encodes a protein of 437 amino acids with a molecular weight of 48 kD and a calculated pI of 5.77. Sequence analysis showed that GhWRI1 contains two AP2 domains and belongs to the AP2 subfamily. Using a green fluorescent protein fusion protein, we demonstrated that GhWRI1 accumulated specifically in the nuclei of onion epidermal cells. Real-time RT-PCR revealed that GhWRI1 was constitutively expressed in true leaves and roots, and its expression in fibers and seeds was upregulated over the course of development. In addition, GhWRI1 enhances the activity of a luciferase reporter by approximately 20-fold, by using a GAL4 activating system in tobacco cells. Our results indicate that GhWRI1 is a strong transcriptional activator. The possible role of cotton GhWRI1 in controlling the seed oil content is discussed.

Key words: Wrinkled1, gene(WRI1, ), Transcription factor, AP2 domain, Transient assay, Gossypium hirsutum

李鑫, 王正明, 薛伟, 初明光. 棉花中一个新型转录因子GhWRI1的表达特征与功能分析[J]. 生物技术通报, 2013, 0(6): 80-86.

Li Xin, Wang Zhengming, Xue Wei, Chu Mingguang. Identification and Characterization of a Novel Gene, GhWRI1, Encoding an AP2-type Transcription Factor in Gossypium hirsutum[J]. Biotechnology Bulletin, 2013, 0(6): 80-86.

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