川草2号老芒麦(Elymus sibiricus L.)atpA基因的克隆及其调控表达  

Molecular Cloning Regulation of Chilling-repressed Gene atpA in Elumus sibiricus

作　　者：何文兴[1] 徐莺[1] 唐琳[1] 魏琴[1] 李静[1] 陈放[1] 

机构地区：[1]四川大学生命科学学院,成都610064

出　　处：《生物化学与生物物理进展》2005年第1期67-74,共8页Progress In Biochemistry and Biophysics

基　　金：国家"十五"科技攻关计划重大项目(2001BA606A-11).~~

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(ElymussibiricusL. 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CF1 α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.ATPase is closely related to chilling tolerance. The EST sequence of chilling-repressed gene atpA encoding CF1 alpha-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 2degreesC 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 4similar to8 h of chilling stress and 16similar to24 h after removing stress. This result provides new clue for revealing the CF1 alpha-subunit role to ATPase in plant defence against chilling stress.

关 键 词：atpA基因 mRNA差异显示(DDRT-PCR) 低温胁迫 川草2号老芒麦 

分 类 号：Q786[生物学—分子生物学]