世界科学技术-中医药现代化2025，Vol.27Issue(1)：56-64,9.DOI:10.11842/wst.20240601003
野菊中三个黄酮苷合成UGT基因克隆及原核表达
Cloning and Prokaryotic Expression of Three Flavonoid Glycoside Synthesizing UGT Genes of Chrysanthemum indicum.L
摘要
Abstract
Objective The three UGT genes involved in the synthesis of flavonoid glycosides in the wild chrysanthemum were cloned,fused and expressed in prokaryotic system,which provided a basis for further study of the functions of these genes in the synthesis of flavonoid compounds.Methods three candidate UDP-glycosyltransferase(UGT)family genes were successfully cloned from flower tissue cDNA of diploid Chrysanthemum indicum.L in Hubei,namely:CiUGT1(1425 bp),CiUGT2(1311 bp)and CiUGT3(1314 bp)encoded opal fraction of 51.72,48.08 and 48.28 kDa,respectively.After the physical and chemical properties of the protein were analyzed,the recombinant plasmid was constructed by seamless Klon technology,and the recombinant proteins of CiUGT2 and CiUGT3 were successfully expressed in Escherichia coli.Results SDS-PAGE analysis showed that the purified CiUGT2 and CiUGT3 protein samples were about 0.2 mg/mL and 1.4 mg/mL,respectively.Conclusion Two candidate UGTs were obtained to catalyze the synthesis of flavonoid 7-O-glucosides in Chrysanthemum indicum.L,which provide the theoretical basis for the molecular mechanism of flavonoid glycosides biosynthesis in the capitulum of Chrysanthemum indicum.L.关键词
野菊/UGT/原核表达/黄酮Key words
Chrysanthemum indicum.L/UGT/Prokaryotic expression/Flavonoid分类
医药卫生引用本文复制引用
秦志伟,廖家豪,张景景,刘义飞,胡志刚,刘晶晶..野菊中三个黄酮苷合成UGT基因克隆及原核表达[J].世界科学技术-中医药现代化,2025,27(1):56-64,9.基金项目
湖北省中医药管理局中医药科研项目(ZY2023F130):湖北两个菊属药典种野菊和菊花(福白菊)品质评价体系构建,负责人:张景景 （ZY2023F130）
湖北省中央引导地方科技发展专项(2022BGE201):中药炮制与饮片等级研究创新服务平台,负责人:胡志刚. （2022BGE201）
