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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2011, Vol. 31 Issue (5): 34-41    
研究报告     
人工microRNA干扰DREB亚族A-5组转录抑制子基因增强了拟南芥对低温和高盐胁迫的耐受性
周露, 董春娟, 刘进元
清华大学生命科学学院 教育部蛋白质科学重点实验室 北京 100084
Increased Resistance of Arabidopsis to Cold and Salt Stresses by Suppresing the Transcription Repressors of the A-5 Group among the DREB Subfamily Transcription Factors through Artificial microRNA
ZHOU Lu, DONG Chun-juan, LIU Jin-yuan
MOE Laboratory of Protein Science, School of Life Science, Tsinghua University, Beijing 100084, China
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摘要:

转录抑制子是一类与DNA的特异位点结合并抑制其附近基因转录的蛋白质,主要通过与转录激活子或基本转录复合物发生作用以及引起染色质重排等3种方式来抑制目标基因的转录。DREB类转录因子的A-5组中共有6个转录抑制子,其功能和作用机制还有待进一步研究。通过设计人工microRNA-A5(amiRNA-A5)使其较特异地作用这些抑制子的mRNA, 并将构建成功的pCAMBIA-pre-amiRNA-A5表达载体转化到拟南芥中,最终得到了9株T2代转基因纯合子。Real-time PCR检测表明,与野生型对照相比,转基因植株中靶基因的表达量有明显降低。抗性分析的结果显示与野生型植株相比转基因植株在低温和高盐胁迫条件下其相对离子渗漏率(REL)和丙二醛(MDA)含量增加都比较少,表明转基因植株对低温和高盐胁迫的耐受性增强,这为进一步研究DREB亚族A-5组转录抑制子在胁迫条件下的功能和作用的分子机制奠定了基础。

关键词: amiRNADREB亚族A-5组转录抑制子基因沉默抗性    
Abstract:

Transcription repressors are proteins that bind to specific sites on DNA and prevent transcription of nearby genes through interacting with transcripitonal activators, acting on the basal transcription factor complex or chromatin remodeling. Among the DREB subfamily, there are 6 transcription repressors containing the EAR motif in the A-5 group, functional analysis of some repressor indictaed it could modulate plant response to stress, however their specific functions and mechanisms are reqiured further research. By designing the amiRNA-A5, which could specifically target the mRNAs of these transcription repressors, and transforming the pCAMBIA-pre-amiRNA-A5 expression vector to Arabiopsis, 9 lines of T2 transgenetic homozygotes were got, selected by Hygromycin and PCR. Real-time PCR assays demonstrated that compared to the wild type plants, the relative expression levels of target genes in transgenetic plants were obviously decreased. To analyze the resistance of these transgenetic plants under cold and salt stresses, relative electrolyte leakage (REL) and malondialdehyde (MDA) content were measured. Both of these two indexes were increased less in the transgenetic plants than in the wilde type plants under stress conditions, suggesting the increased resistance of the transgenetic plants to cold and salt stress. The results indicated these transcription repressors might negatively function under cold and salt stress, provding clues for further functional mechanism study of the transciption repressors of the A-5 group among the DREB subfamily.

Key words: amiRNA    A-5 group of the DREB subfamily    Transcription repressor    Gene silencing    Resistance
收稿日期: 2010-12-23 出版日期: 2011-05-27
ZTFLH:  Q789  
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引用本文:

周露, 董春娟, 刘进元. 人工microRNA干扰DREB亚族A-5组转录抑制子基因增强了拟南芥对低温和高盐胁迫的耐受性[J]. 中国生物工程杂志, 2011, 31(5): 34-41.

ZHOU Lu, DONG Chun-juan, LIU Jin-yuan. Increased Resistance of Arabidopsis to Cold and Salt Stresses by Suppresing the Transcription Repressors of the A-5 Group among the DREB Subfamily Transcription Factors through Artificial microRNA. China Biotechnology, 2011, 31(5): 34-41.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2011/V31/I5/34


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