甘蓝型油菜‘沪油15'中两个CBF类转录因子的克隆与分析  

Cloning and Analysis of CBF Transcription Factors from Brassica napus L.cv.‘Huyou 15'

作　　者：庄静[1,2] 朱波[2] 金晓芬[2] 彭日荷[2] 乔玉山[1] 章镇[1] 熊爱生[2] 姚泉洪[2] 

机构地区：[1]南京农业大学园艺学院,南京210095 [2]上海市农业科学院生物技术研究所,上海201106

出　　处：《植物生理学通讯》2008年第6期1106-1112,共7页Plant Physiology Communications

基　　金：国家“863”项目(2006AA10Z117);上海市自然科学基金(08ZR1417200);上海市青年科技启明星跟踪计划(08QH14021)。

摘　　要：以拟南芥CBF家族转录因子AP2保守域的氨基酸序列为信息探针,搜索油菜UniGene数据库,得到2个UniGene:Bna. 2193和Bna.2260,序列拼接得到2个cDNA序列(BnaCBF-1和BnaCBF-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。序列分析显示,克隆的BnaCBF-1-Hy15与BnaCBF-1基因序列完全一样,而BnaCBF-2-Hy15与BnaCBF-2的基因序列差异很小,只有4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、序列比对、进化树、功能域、二级结构、三级结构进行了预测。结果显示,‘沪油15'来源的BnaCBF-1-Hy15和BnaCBF-2-Hy15转录因子属于AP2/ERF家族中的DREB-A1亚族,是亲水性蛋白,在蛋白质的三级结构上与atCBF1相似。Two CBF family transcriptional regulators were isolated from Brassica napus by in silico cloning method using the conserved domain amino acid sequence of atCBF1 as probe. Based on the sequences of BnaCBF-1 and BnaCBF-2 from Bna.2193 and Bna.2260, BnaCBF-1-Hy15 and BnaCBF-2-Hy15 genes were isolated from B. napus cv. ＇HuyoulS＇ by RT-PCR and PCR using cDNA and DNA as template. DNA sequencing and analysis indicated that there was no difference between BnaCBF-1-Hyl5 and BnaCBF-1, bnt four amino acid residues difference between BnaCBF-2-Hyl5 and BnaCBF-1. No intron localized on the two genes, respectively. Then, cDNA and deduced amino acid sequence, composition, hydrophobicty and hydrophilicity, physical and chemical characterization, phylogenetic tree, conserved domain sequence, function domain, and molecular modeling were predicted and analyzed. BnaCBF-1-Hyl5 and BnaCBF-2-Hyl 5 were part of the DREB-A1 subfamily, and were hydrophilic protein. The two proteins and atCBF1 had similar three-dimension structure.

关 键 词：转录因子 CBF 生物信息学 甘蓝型油菜'沪油15’ 

分 类 号：S565.4[农业科学—作物学]