Cloning and Functional Characterization of a Flavonoids UDP-Glycosyltransferase Gene DsUGT11 from Desmodium Styracifolia
Objective This study aimed to investigate the glycosyltransferase gene DsUGT11 involved in the biosynthesis of flavonoids in Desmodium styracifolia,and to analyze the function of its encoding protein by bioinformatic tools,gene cloning,prokaryotic expression,and other technologies.Methods The sequence characteristics and potential biological functions of DsUGT11 were analyzed and predicted by bioinformatics analysis,respectively.Total RNA was extracted from fresh leaves and reverse transcribed into cDNA,from which DsUGT11 gene was successfully amplified and cloned.Heterologous expressed protein was induced and purified,followed by functional characterization using enzymatic reaction in vitro.Results A candidate glycosyltransferase gene,designated as DsUGT11,was identified from the transcriptome data of D.styracifolia.The length of the open reading frame of DsUGT11 is 1426 bp,and the molecular weight of its encoding protein is expected to be 52.14 KDa.By bioinformatic analysis,DsUGT11 was found to harvest a conserved motif of"PSPG"that is unique to the UGT family.Moreover,DsUGT11 was successfully amplified and cloned using the prokaryotic expression vector pMALc5X.Recombinant protein was induced and purified subsequently.Next,the purified protein was used to perform the enzymatic reaction in vitro,the result of which suggested that DsUGT11 was able to catalyze the conversion of 2-OH-naringenin and UDP-glucose into three different compounds,one of which was authenticated as apigenin-7-O-β-D-glucoside(also known as Apigetrin),with two others unknown.Conclusion In this study,the DsUGT11 gene was identified and cloned,whose encoding protein is a flavone-oxyglycosyltransferase catalyzing the conversion of 2-OH-naringenin and UDP-glucose into three different compounds including Apigetrin.
NETL
NSTL
万方数据
