诸葛菜OvCYP86MF基因的克隆及其特性分析  

CLONING AND MOLECULAR CHARACTERIZATIONS ANALYSES OF CYP450 FROM Orychophrogmus violaceus (L.)

作　　者：王玲平[1,2] 周生茂[3] 曹家树[1] 

机构地区：[1]浙江大学蔬菜研究所,浙江杭州310029 [2]浙江省农业科学院蔬菜研究所,浙江杭州310021 [3]广西农业科学院蔬菜研究所,广西南宁530007

出　　处：《核农学报》2010年第4期728-736,共9页Journal of Nuclear Agricultural Sciences

基　　金：浙江省自然科学基金项目(Y307557)

摘　　要：为进一步阐明细胞色素P450基因CYP86MF在诸葛菜发育中的分子机理,本文根据已知同源基因保守序列设计特异引物,利用RT-PCR和RACE技术从诸葛菜中获得一个细胞色素P450基因(OvCYP86MF)全长cDNA序列,该序列全长为1876bp,含有1605bp的完整开放阅读框,可编码534个氨基酸,分子量和等电点分别为61.4kDa和6.90,具有细胞色素P450蛋白的典型特征,即保守结构域FNAGPRLCIG;原核表达显示该基因的融合蛋白在体外可以诱导表达;DANSTAR和Clustal W软件分析表明该基因的全长cDNA序列及其编码氨基酸序列与十字花科物种拟南芥相似性很高,达到80%以上,亲缘关系最近;与CYP86C亚家族成员在氨基酸水平上的相似性均高于50%,因此推断该基因属于CYP86C这个亚家族。Northern杂交分析表明该基因在花蕾中特异表达。OvCYP86MF,a cytochrome p450 gene,was cloned from Orychophrogmus violaceus（L.） OE Schulz by RT-PCR and RACE techniques.The whole cDNA sequence was 1876bp in length with a open reading frame（ORF） of 1605bp.The ORF encoded 534 amino acid residues,which had a typically conserved domain of cytochrome P450 cysteine heme-iron ligand signatures（FNAGPRLCIG）.The fusion protein was expressed by inducing in vitro.Sequence alignment and Phylogenetic tree analysis showed that OvCYP86MF gene shared more than 80% identities with thoses of Arabidopsis thaliana at the levels of complete cDNA sequences and its deduced amino acid sequences.It also showed over 50% identity to every member of CYP86c family at the level of deduced amino acid sequences.It was inferred that OvCYP86MF belonged to the sub-family CYP86C.Northern hybridization showed that the gene was expressed only in floral buds.

关 键 词：诸葛菜 OvCYP86MF基因 克隆 表达 

分 类 号：S647[农业科学—蔬菜学]