白菜WRKY转录因子cDNA全长的克隆及分析  

Cloning and Characterization of A Full-length cDNA of WRKY Transcription Factor Gene in Brassica campestris ssp.chinensis

作　　者：王彦华[1] 侯喜林[2] 申书兴[1] 

机构地区：[1]河北农业大学园艺学院,保定071001 [2]南京农业大学作物遗传与种质创新国家重点实验室,南京210095

出　　处：《农业生物技术学报》2007年第5期810-815,共6页Journal of Agricultural Biotechnology

基　　金：高等学校博士点科研基金(No.20030307021)资助

摘　　要：参照拟南芥(Arabidopsis thaliana)WRKY转录因子基因保守序列设计引物,利用RT-PCR及RACE技术克隆了白菜(Brassica campestris ssp.chinensis)WRKY转录因子基因cDNA全长,命名为BcWRKY1(GenBank登录号为AY836002)。序列分析发现,白菜BcWRKY1基因核苷酸序列长980bp,推导的氨基酸序列编码285个氨基酸残基,与拟南芥AtWRKY18氨基酸序列的同源性为74%,与拟南芥AtWRKY60同源性为59%;与其它植物WRKY基因的同源性则较低。Southernblot显示该基因在白菜基因组中为单拷贝。半定量RT-PCR分析表明,用2mmol/L的水杨酸处理后2～8h,以及用霜霉病菌(Peronospora parasitica)接种后6～12h,白菜BcWRKY1基因的表达量最高。PCR primers were designed based on the conserved domain of Arabidopsis thaliana WRKY transcription factor. The full-length cDNA of WRKY transcription factor was cloned from Brasssica campestris ssp.chinensis using RT-PCR and rapid amplification cDNA end （RACE） techniques. Sequence analysis indicated that BcWRKY1 gene （GenBank accession number： AY836002） consisted of 980 nucleotides（nt）, and deduced amino acid sequence containing 285 amino acids. Further comparison showed that its identity was 74% and 59% to AtWRKY18 and AtWRKY60 genes of Arabidopsis, respectively. The deduced amino acid sequence of BcWRKY1 gene has lower homology with other plant WRKY genes. Southern blotting analysis indicated there was a single copy in genome of B. campestris ssp.chinensis. Semi-quantitative RT-PCR demonstrated that the corresponding mRNA of BcWRKY1 was accumulated most abundantly 2-8 h after treatment upon 2 mmol/L salicylic acid, and 6-12 h after infection by Peronospora parasitica, respectively.

关 键 词：白菜 转录因子 水杨酸 RACE 半定量RT-PCR 

分 类 号：S188[农业科学—农业基础科学]