受病原细菌诱导表达增强的水稻转录因子基因OsBTF3的分子鉴定  

Molecular Identification and Characterization of A Rice Gene of OsBTF3 Encoding A Transcriptional Factor Up-Regulated by Xanthomonas oryzae pv. oryzae

作　　者：李广旭[1] 吴茂森[1] 吴静[1] 何晨阳[1] 

机构地区：[1]中国农业科学院植物保护研究所/植物病虫害生物学国家重点实验室,北京100193

出　　处：《中国农业科学》2009年第7期2608-2614,共7页Scientia Agricultura Sinica

基　　金：国家重点实验室自主研究课题专项(SKL2007SR06)

摘　　要：【目的】分子克隆和定性分析受水稻白叶枯病菌(Xanthomonas oryzae pv.oryzae,Xoo)诱导的水稻转录因子基因OsBTF3。【方法】用生物信息学和RT-PCR方法鉴定和分离OsBTF3基因全长cDNA序列;用原核表达和亚细胞定位方法分析产物表达及其定位;用RT-Q-PCR方法分析基因的组织表达特性和对Xoo侵染的的应答表达。【结果】从水稻品种日本晴cDNA中克隆了OsBTF3全长序列,它编码具有175个氨基酸的蛋白质,具有核定位信号和NAC保守结构域;OsBTF3与其它植物BTF3序列同源性可达79%～88%;在大肠杆菌中表达产生一个与预测分子量大小一致的27kD蛋白质;OsBTF3::GFP融合蛋白主要在细胞核和细胞膜出现;OsBTF3基因在水稻不同发育期和组织中均有表达,但显著地受Xoo侵染的诱导而增强。【结论】成功分离了一个受Xoo诱导表达增强的水稻转录因子基因OsBTF3;OsBTF3主要定位于细胞核和细胞膜,可能在水稻感病性表达中起调控作用。[ Objective ] To identify and characterize OsBTF3, a rice gene encoding a transcriptional factor up-regulated by Xanthomonas oryzae pv. oryzae （Xoo）. [Method] Identification and isolation of the full length of cDNA sequence of OsBTF3 from rice （Oryza sativa L. cv. Nipponbare） was performed through bioinformatics and RT-PCR analysis. Prokaryotic expression and subcellular localization of OsBTF3 was conducted as well. Expression of OsBTF3 in rice tissues at various developmental stages and in response to Xoo infection was assayed via real-time quantitative PCR （RT-Q-PCR）. [Result] The cloned OsBTF3 gene encodes a putative protein consisting of 175 amino acids with a nuclear localization signal （NLS） in N-terminal and a conserved NAC domain. OsBTF3 shares a high identity with the BTF3 homologues from other species （79%-88%）. Prokaryotic expression analysis shows that an expressed protein （27 kD） is obtained after IPTG induction in E. coli. OsBTF3：：GFP fusion protein mainly appears in the nuclear and cell membrane. RT-Q-PCR analysis indicates that OsBTF3 can express in various tissues of rice plants and be significantly induced by Xoo inoculation. [ Conclusion] The full length cDNA sequence of OsBTF3 is successfully isolated. OsBTF3 is mainly localized in nuclear and cell membrane, and might play an important role in the regulation of susceptibility of rice.

关 键 词：水稻 OsBYF3 基因克隆 表达 感病性 

分 类 号：S511[农业科学—作物学]