关键词: 月季, UGT91A1, 载体构建, 原核表达, pCold-TF, 植物三萜, 生物信息学

Abstract: A glycosyltransferase gene 91A1（UDP glycosyltransferase 91A1，UGT91A1）in Rosa chinensis was mined by phylogenetic tree analysis of triterpenoid glycosyltransferases（GTs） in plants；the physicochemical properties of UGT91A1 were analyzed using bioinformatics methods such as pUGT db，SOPMA，SWISS MODEL，Expasy，WoLF PSORT，TMHMM，etc；pCold‐UGT91A1 recombinant expression vector was constructed using double enzyme digestion method，and protein expression was detected by SDS‐PAGE electrophoresis and Coomassie brilliant blue staining.The results showed that the coding sequence（CDS）of the UGT91A1 gene in Rosa chinensis was 1 491 bp，encoding an extracellular protein containing 493 amino acids，mainly located in the endoplasmic reticulum.The molecular weight was 55. 4 ku and the theoretical isoelectric point was 5.42.Its main sugar donor was uridine diphosphate glucose（UDPG）.The construction of the recombinant expression vector pCold‐UGT91A1 for UGT91A1 and pCold TF was successfully completed.The expressed Trigger Factor‐UGT91A1 fusion protein had a molecular weight of approximately 107 ku，and the HRV3C enzyme could cleave the TF tag（52 ku）in the fusion protein to obtain UGT91A1 protein（55.4 ku）.

Key words: Rosa chinensis, UGT91A1, Carrier construction, Prokaryotic expression, pCold‐TF, Plant triterpenoid, Bioinformatics