福建农业学报2025，Vol.40Issue(8)：757-766,10.DOI:10.19303/j.issn.1008-0384.2025.08.002
斑地锦类黄酮-3'-羟化酶基因的克隆及表达分析
Cloning and Expression Analysis of Flavonoid 3'-hydroxylase Gene in Euphorbia maculata
摘要
Abstract
[Objective]Expressions of flavonoid 3'-hydroxylase gene(F3'H)and promoter in organs of Euphorbia maculata at growth stages were analyzed to study the related biosynthetic pathways in the plant.[Method]Based on the transcriptome sequences,open reading frame(ORF)of F3'H was cloned from E.maculata,and sequence of the gene promoter obtained by Tail-PCR technique with bioinformatics analysis performed.RT-qPCR was applied to determine the expressions in different plant parts at various growth stages.The protein was expressed through prokaryotic system for analysis.[Results]EmF3'H(GenBank accession No.OR944655.1)had an ORF of 1524 bp in length encoded 507 amino acids.It was a hydrophilic transmembrane protein in the cytochrome P450(CYP450)family and subcellularly localized in endoplasmic reticulum.Its 1273 bp promoter contained core elements TATA-box and CAAT-box as well as cis-acting elements related to hormone,stress,and light response.The expressions of EmF3'H significantly differentiated in the organs with the highest level found in the fruits and the lowest in the leaves at the reproductive stage.The target protein of approximately 75 kDa was obtained through prokaryotic expression at the highest level observed 4 h after IPTG induction.[Conclusion]EmF3'H was expressed in various tissues at different growth stages.It played an important role in the biosynthesis of flavonoids in E.maculata.关键词
斑地锦/类黄酮-3'-羟化酶/RT-qPCR/原核表达/槲皮素Key words
Euphorbia maculata/flavonoid 3'-hydroxylase/RT-qPCR/prokaryotic expression/quercetin分类
生物学引用本文复制引用
桂明明,于武华,朱婧姝,叶菲,陈春香,黄胜和,饶泽昌..斑地锦类黄酮-3'-羟化酶基因的克隆及表达分析[J].福建农业学报,2025,40(8):757-766,10.基金项目
国家自然科学基金项目(32260926) （32260926）
江西省教育厅科学技术研究项目(GJJ2406003、GJJ213308) （GJJ2406003、GJJ213308）
江西中医药高等专科学校科研项目(2024XJKY01) （2024XJKY01）
江西中医药高等专科学校科研创新团队项目(2022CX01) （2022CX01）
