沪油15中两个AP2/ERF-B1亚族转录因子的克隆与分析  

CLONING AND ANALYSIS OF AP2/ERF-B1 SUBFAMILY TRANSCRIPTION FACTORS FROM Brassica napus L. HUYOU 15

作　　者：庄静[1,2] 彭日荷[1] 高峰[1] 付晓燕[1] 朱波[1] 金晓芬[1] Jian Zhang 熊爱生[1] 姚泉洪[1] 

机构地区：[1]上海市农业科学院,上海201106 [2]加拿大艾伯塔省研究院,埃德蒙顿T9C1T4

出　　处：《核农学报》2009年第3期435-441,453,共8页Journal of Nuclear Agricultural Sciences

基　　金：国家863项目(2006AA10Z117;2008AA10Z401);上海市青年科技启明星跟踪计划(08QH14021);上海市自然科学基金(08ZR1417200);上海市国际合作项目(08540706500)

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。Two AP2/ERF family transcription factors （BnaERFBI-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 BnaERFB1-1-Hy15, BnaERFB1-2 and BnaERFB1-2-Hy15 and both no intron localized on the two genes from HuyoulS, 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 threedimension structure, and BnaERFB1-1 was mainly expression in seed, while BnaERFB1-2 was mainly expression in vegetative meristem.

关 键 词：AP2/ERF B1亚族 转录因子 克隆 沪油15 

分 类 号：S565.4[农业科学—作物学]