中国现代中药2025，Vol.27Issue(11)：2038-2046,9.DOI:10.13313/j.issn.1673-4890.20250314009
广西莪术萜类合酶基因CkTPS及其启动子克隆与表达分析
Cloning and Expression Analysis of Terpene Synthase CkTPS Gene and Promoter from Curcuma kwangsiensis
摘要
Abstract
Objective:To clone a terpene synthase(TPS)gene and its promoter sequence from Curcuma kwangsiensis S.G.Lee et C.F.Liang and analyze their expression patterens.Methods:Reverse transcription-polymerase chain reaction(RT-PCR)was used to clone the TPS gene from Curcuma kwangsiensis(CkTPS gene)and its promoter sequence.Various bioinformatics tools were employed to analyze and predict physicochemical properties,protein structure,conserved domains,and promoter cis-acting elements.The tissue-specific expression patterns of the CkTPS gene at different developmental stages and under exogenous methyl jasmonate(MeJA)treatment were analyzed using quantitative real-time PCR(qRT-PCR).Results:The open reading frame of CkTPS is 1 650 bp,encoding 549 amino acids.With a molecular weight of 63.72 kDa,a theoretical isoelectric point of 5.25,and an aliphatic index of 94.46,CkTPS is an unstable,non-transmembrane,and hydrophilic protein.The CkTPS protein contains the conserved motifs DDXD and NSE/DTE.It shows 99%homology with TPS proteins from Curcuma wenyujin,Curcuma zedoaria,Zingiber officinale,and Zingiber zerumbet,clustering in the same branch of the phylogenetic tree.The cloned promoter is 1 807 bp in length and contains cis-acting elements,including TATA-box,CAAT-box,CGTCA-motif,and G-box.The results indicate that CkTPS shows significant differences in relative expression among different tissues at various developmental stages of Curcuma kwangsiensis,with higher expression in tubers and rhizomes than in leaves and petioles.After MeJA treatment,CkTPS expression is significantly upregulated in different tissues.Conclusion:The CkTPS gene is involved in the regulation of terpenoid biosynthesis in Curcuma kwangsiensis,which may be induced by MeJA.关键词
广西莪术/萜类合酶/生物信息学分析/启动子元件分析/表达分析Key words
Curcuma kwangsiensis S.G.Lee et C.F.Liang/terpene synthase/bioinformatics analysis/promoter element analysis/expression analysis分类
医药卫生引用本文复制引用
谢月英,姚李祥,卢海,熊峥,潘春柳,黄雪彦,余丽莹,周芸伊..广西莪术萜类合酶基因CkTPS及其启动子克隆与表达分析[J].中国现代中药,2025,27(11):2038-2046,9.基金项目
广西自然科学基金项目(2022GXNSFBA035631,2024GXNSFAA010196) （2022GXNSFBA035631,2024GXNSFAA010196）
广西药用植物园科研基金项目(GYJ202104) （GYJ202104）
广西青年岐黄学者培养项目(GXQH202411) （GXQH202411）
中央本级重大增减支项目(2060302) （2060302）
