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

摘要： 通过植物三萜类糖基转移酶（GTs）的系统进化树分析挖掘出一个月季糖基转移酶基因91A1（UDP‐glycosyltransferase 91A1，UGT91A1）；采用pUGTdb、SOPMA、SWISS MODEL、Expasy、WoLF PSORT、TMHMM等生物信息学方法分析UGT91A1的理化性质；利用双酶切法构建pCold-UGT91A1重组表达载体，SDS-PAGE电泳和考马斯亮蓝染色检测蛋白质表达。结果表明，月季UGT91A1基因的编码序列（CDS）为1 491 bp，编码一个含493个氨基酸的跨膜蛋白，其主要定位于内质网，分子质量为55.4 ku，理论等电点为5.42，作用的糖基供体主要为尿苷二磷酸葡萄糖（UDPG）。成功构建了UGT91A1 和pCold-TF 的重组表达载体pCold-UGT91A1，其表达的Trigger Factor-UGT91A1 融合蛋白分子质量为107 ku，
HRV3C酶切该融合蛋白得到去除TF标签（52 ku）的UGT91A1蛋白（55.4 ku）。

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