十字花科植物CYP86MF同源基因的克隆及特征分析  

Cloning and Identification of CYP86MF Homologous Genes from Cruciferae

作　　者：王玲平[1,2] 曹家树[1] 叶纨芝[1] 向珣[1] 余小林[1] 

机构地区：[1]浙江大学蔬菜研究所 [2]浙江省农科院蔬菜研究所,杭州310021

出　　处：《农业生物技术学报》2005年第2期145-151,共7页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.30370975);浙江省重大科技项目(No.021102536)资助

摘　　要：用保守的特异引物进行PCR扩增,并结合RACE技术,从大白菜(BrassicacampestrisL.)品种黄芽14,萝卜(RaphanussativusL.)品种圆白和诸葛菜(Orychophrogmusviolaceus)3个材料中扩增到3个完整的cDNA全长,分别命名为Bc-CYP86MF,RsCYP86MF和OvCYP86MF,它们的cDNA全长分别为1854,1818和1876bp,分别编码524,526和534个氨基酸残基。将所推导的氨基酸序列与数据库中已知基因进行排序,发现它们与拟南芥CYP86家族所有成员的氨基酸相似性都大于38%,可将它们归为细胞色素CYP86基因家族。BcCYP86MF和RsCYP86MF与CYP86C4亚家族氨基酸同源性均大于55%,则可归为CYP86C4亚家族,而OvCYP86MF与CYP86C3的同源性最高,为86.5%,应归为CYP86C3亚家族。根据氨基酸序列的排序结果构建的系统树表明,CYP450基因的氨基酸序列进化程度与物种的亲缘关系的远近相吻合。对蛋白二级结构预测结果发现,它们都具有细胞色素P450典型的保守结构域FNAGPRLCIG,而且氨基酸组成也很相似。Three full-length cDNA sequences which were 1 854, 1 818 and 1 876 bp in length, named BcCYP86MF, RsCYP86MF and OvCYP86MF, and contained open reading frames of 524, 526 and 534 amino acid residues, respectively were cloned from Brassica campestris L. ssp. pekinensis Olsson. cv. Huangya-14, Raphanus sativus L.cv. Yuanbai and Orychophrogmus violaceus (L.) OE Schulz by SMART RACE RT-PCR strategy with gene specific primers. They were classified into CYP86 family for over 38% identities of the amino acid sequences of the three genes to CYP86 genes from Arabidopsis thalia, whereas BcCYP86MF and RsCYP86MF were sorted into CYP86C4 subfamily in terms of over 55% similarities to CYP86C4 at the amino acid level, and OvCYP86MF into CYP86C3 subfamily according to 86.5% homology between them at the amino acid level. The phylogenetic tree showed that evolution of amino acid sequences deduced from the CYP450 genes was basically in accordance with the phylogenetic relationship of these plants. They had similar amino acid component and contained a conserved domain of cytochrome P450 cysteine heme-iron ligand signature (FNAGPRLCIG).

关 键 词：十字花科植物 特征分析 同源基因 细胞色素P450 氨基酸序列 克隆 RACE技术 氨基酸同源性 P450基因 蛋白二级结构 cDNA PCR扩增 氨基酸残基 保守结构域 氨基酸组成 特异引物 基因家族 亲缘关系 预测结果 大白菜 诸葛菜 数据库 

分 类 号：Q949.748.3[生物学—植物学] Q78