川草 2 号老芒麦 (Elymus sibiricus L.) atpA 基因的克隆及其调控表达
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国家“十五”科技攻关计划重大项目(2001BA606A-11).


Molecular Cloning Regulation of Chilling-repressed Gene atpA in Elumus sibiricus
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This work was supported by The National Key Sciences and Technology R&D Program in the 10th Five-year Plan of China (2001BA606A-11).

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    摘要:

    ATPase 与植物的耐寒性密切相关 . 运用 mRNA 差异显示技术 (DDRT-PCR) 从耐寒植物川草 2 号老芒麦 (Elymus sibiricus L. cv. ‘ chuancao No.2 ' ) 中获得了受冷抑制的 atpA 基因表达序列标签 (EST) 序列,通过 5 ′ cDNA 末端快速扩增 (RACE) 获得了 atpA 基因全长 cDNA 为 1 754 bp ,开放阅读框 (ORF) 长 1 518 bp ,编码 505 个氨基酸 . 氨基酸序列与小麦、水稻、玉米 atpA 基因分别具有 95 %、 94 %、 94 %的相似性 . 对 2 ℃低温胁迫及解除胁迫后恢复过程中共 13 个时段的 RNA 印迹分析表明, atpA 基因在低温胁迫 12 h 转录受到强烈抑制,在低温胁迫开始后的 4 ~ 8 h 及解除胁迫后的 16 ~ 24 h ,其转录水平明显强于对照 . 实验结果为揭示 CF1α亚基对调控 ATPase 在植物御冷性反应的作用,以及α亚基在植物低温信号转导中的作用提供了新的线索 .

    Abstract:

    ATPase is closely related to chilling tolerance. The EST sequence of chilling-repressed gene atpA encoding CF1 α -subunit was obtained from Elumus sibiricus L. cv.'chuancao No.2' by reverse transcription-polymerase chain reactiom (RT-PCR) differential display. Full-length cDNA of 1 754 bp was cloned by 5 ′ RACE. The atpA has an open reading frame (ORF) of 1 518 bp that encodes a precursor protein of 505 amino acid residues. The deduced amino acid sequence exhibits 95%, 94% and 94% positional identity with atpA of wheat, rice and corn, respectively. Northern hybridization analysis on mRNA from treatment at 2 ℃ and post-treatment, in total of 13 time stages, showed that its RNA transcript was strongly inhibited after 12 h of chilling stress, whereas it was clearly higher than control during 4~8 h of chilling stress and 16~24 h after removing stress. This result provides new clue for revealing the CF1α -subunit role to ATPase in plant defence against chilling stress

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何文兴,徐 莺,唐 琳,魏 琴,李 静,陈 放.川草 2 号老芒麦 (Elymus sibiricus L.) atpA 基因的克隆及其调控表达[J].生物化学与生物物理进展,2005,32(1):67-74

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