甘蓝型油菜沪油15中BnaRAV-2-HY15转录因子的克隆及表达分析

CLONING AND EXPRESSION ANALYSIS OF TRANSCRIPTION FACTOR BnaRAV-2-HY15 FROM Brassica napus L. HUYOU 15

基于拟南芥RAV类转录因子的序列,通过电子克隆的方法获得甘蓝型油菜的BnaRAV-2基因序列,再利用RT-PCR方法从甘蓝型油菜沪油15中扩增出编码RAV类转录因子基因BnaRAV-2-HY15,并进行了序列和进化分析。结果显示,该转录因子基因长1119 bp,编码372个氨基酸,含有相对保守的AP2结合域和B3结构域,具有典型的植物RAV类转录因子的结构特征。BnaRAV-2-HY15和AtRAV2具有相似的三维结构。采用半定量RT-PCR方法对基因表达水平进行分析,发现BnaRAV-2-HY15受到PEG、高盐和低温的诱导表达。BnaRAV-2-HY15基因在沪油15盛花和籽粒发育时期的根、茎、叶、花、蕾和荚中均检测不到明显的表达。

The sequences of BnaRAV-2 gene from Brassica napus were obtained by in silico cloning method based on the RAV factor sequence of Arabidopsis,One of RAV family genes,BnaRAV-2-HY15,was isolated by RT-PCR method from B.napus L.cv Huyou15.Then,cDNA and deduced amino acid sequence,phylogenetic tree,and molecular modeling were predicted and analyzed.The coding region of the cloned gene was 1119 bp in length,encoding a 372 amino acids protein.BnaRAV-1-HY15 contained two distinct DNA-binding domains mainly found in higher plants RAV family factors,one AP2 domain together with one B3 domain.BnaRAV-2-HY15 and AtRAV2 had similar three-dimension structure.The results of semi-quantitative RT-PCR analysis for Huyou15 exposed to various treatments demonstrated that the mRNA of gene BnaRAV-2-HY15 was induced by PEG,high-salinity and low-temperature stresses.The gene expression was not detected from the tissues of pod,bud,petal,leaf,stem and root of normally grown Huyou15 plant at period of flowering and seed development.