世界科学技术-中医药现代化2025，Vol.27Issue(10)：2800-2809,10.DOI:10.11842/wst.20250106005
知母糖基转移酶UGT708Z1基因的克隆与功能分析
Cloning and Expression Analysis of a Glycosyltransferase UGT708Z1 Gene from Anemarrhena asphodeloides
摘要
Abstract
Objective To clone the glycosyltransferase gene UGT708Z1 in Anemarrhena asphodeloides and perform its bioinformatics analysis,prokaryotic expression analysis and functional characterization.Methods A candidate glycosyltransferase gene UGT708Z1 was mined and screened out from Anemarrhena asphodeloides based on the transcriptome data.According to its full-length open reading frame,the specific primers with homologous arms were designed.Subsequently,the UGT708Z1 gene was cloned by PCR.The prokaryotic expression recombinant vector pET-32a(+)-UGT708Z1 was constructed through homologous recombination technology,and the soluble target protein was obtained by prokaryotic expression and purified protein technology.Finally,the function of UGT708Z1 was identified through enzymatic reaction in vitro.Results Sequence analysis showed that the open reading frame of UGT708Z1 was 1377 bp,encoding 458 amino acids.The result of prokaryotic expression showed that UGT708Z1 successfully expressed the soluble target protein,and the purified recombinant protein was 70.86 kDa.The results of enzymatic reaction in vitro showed that UGT708Z1 had flavonoid 7-OH glycosylation activity and could catalyze icaritin to produce icariside I.In addition,UGT708Z1 also possessed the activities of catalyzing the 7-O-glycosylation of quercetin and apigenin.Conclusion In this study,a flavonol glycosyltransferase UGT708Z1 was successfully cloned and identified from Anemarrhena asphodeloides,which would lay a foundation for further analysis of flavonol glycosides biosynthesis.关键词
知母/糖基转移酶/基因克隆/原核表达/功能表征Key words
Anemarrhena asphodeloides/Glycosyltransferase/Gene cloning/Prokaryotic expression/Functional characterization分类
生物科学引用本文复制引用
张倩,纪中菊,李志新,董子舒,刘红宁,王晓云,黄佳..知母糖基转移酶UGT708Z1基因的克隆与功能分析[J].世界科学技术-中医药现代化,2025,27(10):2800-2809,10.基金项目
江西省科学技术厅青年基金项目(20242BAB20452):中药活性成分知母皂苷BII生物合成的糖基化分子机制研究 （20242BAB20452）
负责人:黄佳. （）
