A novel cytochrome P450 gene from Catharanthus roseus cell line C20hi: cloning and characterization of expression
- 作者：Lihong He
- 关键词：T16H (CYP71D12) ; tabersonine 16-hydroxylase ; STR ; strictosidine synthase ; D4H ; desacetoxyvindoline 4-hydroxylase ; SSH ; subtractive suppressed hybridization ; EST ; expressed sequence tag ; 2 ; 4-DCAA ; 2 ; 4-dichlorophenoxy acetic acid ; KT ; kinetin ; RT-PCR ; rev
- 刊名：Acta Pharmaceutica Sinica B
- 出版年：2012
- 期刊代码：pca379_22113835
- 类别：pha
- 出版时间：June, 2012
- 卷：2
- 期：3
- 页码：250-255
- 文件大小：635 K
摘要
An expressed sequence tag (EST) obtained from a subtractive-suppression hybridization cDNA library constructed using Catharanthus roseus cell line C20hi and its parental cell line C20D was used to clone a full-length cytochrome P450 cDNA of cyp71d1. The encoded polypeptide contained 507 amino acids with 39-56%identity to other CYP71D subfamily members at the amino acid level. Expression characteristics of cyp71d1 were determined using semi-quantitative RT-PCR. The cyp71d1 transcript was expressed in all three cell lines with the highest level in the cell line C20hi. In the mature C. roseus plant, the cyp71d1 cDNA was highly expressed in petals, roots and stems, but very weakly expressed in young leaves. Its transcription level increased with the development of flowers. 2,4-D could down-regulate the transcription of cyp71d1, as did KT, but only to a minor degree. Neither light nor yeast elicitor could induce the transcription of cyp71d1.