中药材2025，Vol.48Issue(4)：843-847,5.DOI:10.13863/j.issn1001-4454.2025.04.007
鸢尾糖基转移酶基因ItUGT476克隆及原核表达分析
Cloning and Prokaryotic Expression Analysis of Glycosyltransferase Gene ItUGT476 from Iris tectorum
摘要
Abstract
Objective:To clone the glycosyltransferase ItUGT476 gene from Iris tectorum and to perform bioinformatics,tissue ex-pression and prokaryotic expression informations.Methods:The specific primers were designed with the ItUGT476 gene sequence in Iris tectorum transcriptome and amplified by PCR.The products were sequenced and bioinformatic analysis was performed.The expression of ItUGT476 gene in plant tissues was detected by real-time fluorescence quantitative PCR.The recombinant vector pET-32a(+)-ItUGT476 was constructed by homologous recombination technique to express the protein in Escherichia coli.Results:The length of open reading frame of ItUGT476 gene was 1 437 bp,encoding 478 amino acids.The expression level of ItUGT476 in different tissues was rhi-zome,leaf,flower.Evolutionary tree analysis showed that ItUGT476 encoded protein and known flavonoid UGTs clustered on the 7-O-UGT branch.Prokaryotic expression showed that ItUGT476 gene expressed 72.10 kDa target protein.Conclusion:In this study,glycosyl-transferase gene ItUGT476 is cloned from Iris tectorum,which lays the necessary research foundation for the subsequent functional char-acterization of this gene and the analysis of the mechanism of Iris tectorum flavonoid biosynthesis pathway.关键词
鸢尾/糖基转移酶/基因克隆/蛋白原核表达/实时荧光定量PCRKey words
Iris tectorum Maxim./Glycosyltransferase/Gene cloning/Prokaryotic expression of protein/Real-time fluorescence quantitative PCR分类
医药卫生引用本文复制引用
周娇娇,李静,张倩,明良山,樊启猛,季爱加,黄佳..鸢尾糖基转移酶基因ItUGT476克隆及原核表达分析[J].中药材,2025,48(4):843-847,5.基金项目
江西省教育厅科学技术研究项目青年项目(GJJ2200973) （GJJ2200973）
江西省药物分子设计与评价重点实验室开放基金资助项目(JKLDE-KF-2301) （JKLDE-KF-2301）
江西中医药大学博士科研启动基金项目(2022BSZR001) （2022BSZR001）
江西省卫生健康委员会科技计划项目(202311138) （202311138）
