紫茎泽兰CYP75基因cDNA片段的克隆与鉴定

根据菊科植物P450基因CYP75 B5(GenEMBL AF313489)和C YP75 B6(GenEMBL AF313488)核酸序列同源区设计引物,用RT-PCR方法从紫茎泽兰植株中获得一大小为378bp的细胞色素P450基因片段。经克隆、测序及氨基酸序列同源性比较,发现由该片段推导出的氨基酸序列与C YP75 B5和C YP75 B6的氨基酸序列分别具有79.5%和85.6%的同源性,与C YP76 B1、C YP81 B1 v1、C YP81 E1 v2同源性分别为40.8%、35.2%、35.0%,与C YP73 A家族的同源性在28.9%～31.4%;所绘制的系谱树与同源性分析结果一致。因此,初步确定该序列为C YP75家族中某一成员的结构基因片段。

Cloning and identification of a cDNA fragment of CYP75 gene from Eupatorium adenophorum

Based on the homologous nucleotide sequences of CYP75B5(GenEMBL AF313489)and CYP75B6(GenEMBL AF313488)of Callistephus chinensis,a pair of primers were designed and used to amplify the cytochrome P450 gene from the total RNA of Eupatorium adenophorum by RT-PCR,and a cDNA fragment of 378bp in length was obtained.A positive clone was selected for sequencing.The identity between the cloned sequence and CYP75B5,CYP75B6,CPY76B1,CYP81B1v1 and CYP81E1v2 was 79.5%, 85.6%,40.8%,35.2% and 35.0%,respectively,whereas the identity between the cloned sequence and the members of CYP73A varied from 28.9% to 31.4%.The constructed dendrogram showed a similar result to the homologous analysis.Thus the cloned sequence could primarily classified as a member of the CYP75 family.