中国现代中药2026，Vol.28Issue(7)：1334-1342,中插7-中插9,12.DOI:10.13313/j.issn.1673-4890.20260417006
斑地锦3-脱氢奎尼酸脱水酶基因(EmDHQD)及其启动子克隆与表达分析
Cloning and Expression Analysis of 3-Dehydroquinate Dehydratase Gene EmDHQD and Its Promoter in Euphorbia maculata
摘要
Abstract
Objective:To clone the 3-dehydroquinate dehydratase(DHQD)gene EmDHQD and its promoter of Euphorbia maculata and analyze the expression patterns of this gene in different tissue samples at various growth stages.Methods:PCR was employed to clone EmDHQD and its promoter from E.maculata,followed by bioinformatics analysis.The expression patterns of EmDHQD in different tissue samples at different growth stages were examined by real-time quantitative PCR.Results:The open reading frame(ORF)of EmDHQD(GenBank accession No.PX984471)was amplified,with a length of 1 596 bp,encoding 531 amino acid residues.The predicted molecular mass of the protein was 57.34 kDa.The protein was predicted to localized to the mitochondria and belonged to the PLN02520 family.It was hypothesized that EmDHQD might be a soluble enzyme protein stably present in the cytoplasm.The amino acid sequence of EmDHQD shared 79.32%-93.22%similarity with those of DHQD proteins from 21 other plant species,with the highest similarity to that of the DHQD protein of Euphorbia lathyris.A promoter fragment of the EmDHQD gene,approximately 1 700 bp in length,was cloned.In addition to the core eukaryotic promoter elements(TATA-box and CAAT-box),the promoter harbored cis-acting elements responsive to plant hormones,defenses,stress,and low temperatures.EmDHQD was expressed in both stems and leaves during the reproductive growth stage.The relative expression level of EmDHQD in stems was higher than that in leaves on days 1,30,45,and 60 after the first flower opened(P<0.01),while on day 15 after the first flower opened,the relative expression in stems was lower than that in leaves(P<0.01).These results indicated that EmDHQD expression exhibited temporal and tissue specificity.Conclusion:The EmDHQD gene and its promoter are successfully cloned and preliminarily characterized,which provides a theoretical reference for subsequent research on the regulation of gallic acid biosynthesis by EmDHQD in E.maculata.关键词
斑地锦/3-脱氢奎尼酸脱水酶/基因克隆/生物信息学分析/表达分析Key words
Euphorbia maculata L./DHQD/gene cloning/bioinformatics analysis/expression analysis分类
医药卫生引用本文复制引用
乔继琛,桂明明,崔令花,宋美玲,郭三保,于武华,吴晴阳,黄胜和..斑地锦3-脱氢奎尼酸脱水酶基因(EmDHQD)及其启动子克隆与表达分析[J].中国现代中药,2026,28(7):1334-1342,中插7-中插9,12.基金项目
江西省教育厅科学技术研究项目(GJJ2406001) （GJJ2406001）
江西中医药高等专科学校旴江医派研究院科研项目(2024XJKY01) （2024XJKY01）
