【摘要】 利用油菜EST数据库,以拟南芥ERF-B1亚族转录因子序列为探针,电子克隆拼接得到2个cDNA序列(BnaERFB1-1和BnaERFB1-2),通过PCR和RT-PCR方法分别从甘蓝型油菜沪油15的DNA和cDNA中克隆了上述基因。克隆转录因子BnaERFB1-1-Hy15和BnaERFB1-2-Hy15与电子克隆的序列差异很小,分别只有2个和6个氨基酸位点不同,且都没有内含子。从cDNA序列、氨基酸序列的相似性、组成成分、理化性质、疏水性/亲水性、进化树、序列比对、功能域、二级和三级结构等方面进行了分析,结果显示,BnaERFB1-1-Hy15和BnaERFB1-2-Hy15转录因子属于AP2/ERF家族中的ERF-B1亚族,是亲水性蛋白,在蛋白质的三级结构上与拟南芥atERF7相似。EST丰度分析显示,BnaERFB1-1的表达主要集中在种子中,而BnaERFB1-2的表达则主要集中在分生组织中。更多还原

【Abstract】 Two AP2/ERF family transcription factors(BnaERFB1-1 and BnaERFB1-2) were isolated from Brassica napus by in silico cloning method using conserved domain amino acid sequence of Arabidopsis thaliana ERF-B1 subfamily as probe.Based on the sequences of BnaERFB1-1 and BnaERFB1-2,BnaERFB1-1-Hy15,and BnaERFB1-2-Hy15 genes were isolated from B.napus L.Huyou15 by PCR and RT-PCR using DNA and cDNA as template.DNA sequencing and analyses indicated that there were only two and six amino acid residues difference between BnaERFB1-1 and BnaERFB11Hy15,BnaERFB1-2 and BnaERFB1-2-Hy15 and both no intron localized on the two genes from Huyou15,respectively.Then,cDNA and deduced amino acid sequence,composition,physical and chemical characterization,hydrophobicty/hydrophilicity,phylogenetic tree,conserved domain sequences,function domain and molecular modeling were analyzed.The results showed that BnaERFB1-1-Hy15 and BnaERFB1-2-Hy15 were hydrophilic protein,the two proteins and atERF7 had similar three-dimension structure,and BnaERFB1-1 was mainly expression in seed,while BnaERFB12 was mainly expression in vegetative meristem.更多还原