水稻TAF12b基因cDNA克隆及其分子特性鉴定  

cDNA Cloning and Molecular Characterization of OsTAF12b Gene in Oryza sativa

作　　者：齐盼盼 郭留明 李静[2] 吕明芳[2] 袁正杰[2] 张恒木 QI Panpan;GUO Liuming;LI Jing;LÜMingfang;YUAN Zhengjie;ZHANG Hengmu(College of Chemistry and Life Science,Zhejiang Normal University,Jinhua 321004,China;Institute of Virology and Biotechnology,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China)

机构地区：[1]浙江师范大学化学与生命科学学院,浙江金华321004 [2]浙江省农业科学院病毒学与生物技术研究所,浙江杭州310021

出　　处：《中国水稻科学》2023年第6期577-586,共10页Chinese Journal of Rice Science

基　　金：国家自然科学基金资助项目(31972980,2001869);浙江省万人计划资助项目(2019R52033)。

摘　　要：【目的】明确水稻通用转录因子TFⅡD复合物组分中的OsTAF12b的选择性剪接形式并鉴定其亚细胞定位及其表达模式,为深入研究OsTAF12b功能提供基础性信息。【方法】利用5'-/3'-RACE技术扩增并克隆了OsTAF12b基因的全长cDNA;通过生物信息学进行了多重序列比对和进化树构建;利用激光共聚焦显微镜观察OsTAF12b的亚细胞定位;通过qRT-PCR技术分析了该基因在非生物逆境下的表达模式。【结果】发现OsTAF12b基因有4个选择性剪接转录本,其在编码区内仅存在一个赖氨酸的差异。OsTAF12b与其他禾本科植物成员高度同源且在进化树中聚在一个分支上。在本氏烟叶片细胞和水稻原生质体中融合GFP标签的OsTAF12b蛋白均与细胞核标记蛋白H2B共定位。OsTAF12b转录本在水稻叶片中的积累水平较高,而且在多种非生物逆境胁迫下显著上调表达。【结论】水稻OsTAF12b基因存在4种选择性剪接产物,可编码2个仅相差1个赖氨酸残基的细胞核蛋白。表达模式分析表明OsTAF12b可能参与水稻多种非生物逆境胁迫响应过程。【Objective】In order to determine the alternative splicing forms of TATA-box binding protein(TBP)associated factor 12b(OsTAF12b),a component of the general transcription factorⅡD(TFⅡD),in rice(Oryza sativa),characterize their subcellular localization and expression patterns,and provide basal information for further functional studies.【Method】The full-length cDNA of OsTAF12b was amplified and cloned by rapid amplification of cDNA 5’-/3’-ends(RACE).Multiple sequence alignment and a phylogenetic tree were conducted by bioinformatic analysis.The subcellular localization of OsTAF12b was observed with a laser confocal microscopy and its expression patterns under abiotic stresses were analyzed by qRT-PCR.【Results】Four alternative splicing forms of OsTAF12b were identified and there was only one lysine difference within their coding regions.OsTAF12b was highly homologous to those of other gramineous plants and they were grouped together in a clade in the phylogenetic tree.The GFP-fused OsTAF12b protein was colocalized with H2B,a marker labeling nuclear localization,in both cells of N.benthamiana leaves and rice protoplasts.Its transcript level was higher in leaves and significantly up-regulated when exposed to different abiotic stresses.【Conclusion】The OsTAF12b gene has four alternative splicing forms of transcripts and encodes two nuclear proteins with only a single lysine residue difference.Its expression patterns suggest that OsTAF12 could be involved in response to a variety of abiotic stresses.

关 键 词：TAF12b RACE 亚细胞定位 非生物胁迫 表达模式 

分 类 号：S511[农业科学—作物学]