摘要
目的对续断中UGT基因进行全基因组鉴定、组学联合分析及原核表达,以期解析续断三萜皂苷的生物合成。方法采用实验室自测的续断基因组蛋白序列文件,利用BLASTP和hmmsearch,对续断UGT基因序列保守域进行验证。采用Prot-Param、SOMPA、MAGA7.0、Tbtools等工具,对续断UGT基因家族进行蛋白理化性质、蛋白结构、共线性分析等进行分析,并利用转录组数据和代谢组学数据联合分析,筛选出两条可能与三萜皂苷生物合成相关的糖基转移酶,构建其表达载体进行原核表达。结果本研究从续断基因组中鉴定出44个续断UGT基因,续断UGT蛋白长度为49-1083氨基酸,平均分子量为24.86 kDa,等电点为4.31-8.59,续断UGT基因家族共分布在8条染色体上。根据续断、拟南芥和已经鉴定的UGT序列构建的系统发育树表明,续断中的糖基转移酶基因家族主要分布在UGT73、UGT81、UGT85、UGT80家族。顺式作用元件分析表明,光响应元件是UGT基因家族成员启动子区域最普遍存在的元件。利用转录组学和代谢组学联合分析筛选出两条可能与三萜皂苷生物合成相关的糖基转移酶,成功进行了原核表达。结论本研究利用多组学联合筛选出与续断三萜皂苷生物合成相关的两个候选基因进行原核表达,为进一步验证该基因的功能提供参考。
Objective To explore the biosynthesis of Dipsacus asper Wall.ex Henry triterpenoid saponin,and the UGT gene in Dipsacus asper Wall.ex Henry has been analyzed by the identification of whole genome,genome and prokaryotic expression.Methods The laboratory self-tested sequenced protein sequence files of the Dipsacus asper Wall.ex Henry genome were used.To validate the conserved domains of the sequence of the Dipsacus asper Wall.ex HenryUGT gene,BLASTP and hmmsearch were utilized.Prot-Param,SOMPA,MAGA7.0,Tbtools and other tools were used to investigate the protein physicochemical properties,protein structure,and covariance analysis of the Dipsacus asper Wall.ex Henry UGT gene family,and using the joint analysis of transcriptomic data and metabolomics data,two glycosyltransferases that might be related to triterpene saponin biosynthesis were screened,and expression vectors were constructed for prokaryotic expression.Results 44 Dipsacus asper Wall.ex Henry UGT genes were identified from the Dipsacus asper Wall.ex Henry genome.The length of Dipsacus asper Wall.ex Henry UGT proteins ranged from 49 to 1083 amino acids,with an average molecular weight of 24.86 kDa and an isoelectric point of 4.31-8.59.Dipsacus asper Wall.ex Henry UGT gene family was distributed on eight chromosomes.The phylogenetic tree constructed from the sequences of Dipsacus asper Wall.ex Henry,Arabidopsis thaliana and identified UGTs showed that glycosyltransferase gene families in Dipsacus asper Wall.ex Henry were mainly in the UGT73,UGT81,UGT85,and UGT80 families.Cis-acting element analysis showed that light-responsive elements were the most prevalent elements in the promoter regions of UGT gene family members.Two glycosyltransferases potentially related to triterpene saponin biosynthesis were screened using combined transcriptomics and metabolomics analysis,and were successfully expressed in prokaryotic form.Conclusion In this study,two candidate genes related to the biosynthesis of Dipsacus asper Wall.ex Henry triterpenoid saponins were jointly screened for prokaryotic expression using multi-omics,and were subjected to prokaryotic expression for further validation of the function of the genes.
作者
田梅
尹彦棚
王双宜
朱泽宇
谭友莉
侯飞侠
高继海
TIAN Mei;YIN Yanpeng;WANG Shuangyi;ZHU Zeyu;TAN Youli;HOU Feixia;GAO Jihai(State Key Laboratory of Southwest Chinese Medicine Resources,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Sport Hospital Affiliated to Chengdu Sport University,Chengdu 610041,China)
出处
《世界科学技术-中医药现代化》
北大核心
2025年第7期2035-2049,共15页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家中医药多学科交叉创新团队项目:西南特色中药资源多维评价多学科交叉创新团队(ZYYCXTD-D-202209),负责人:彭成
成都体育学院附属体育医院2022年度临床创新课题(LCCX22B02),负责人:谭友莉。
关键词
续断
三萜皂苷
UGT
基因表达分析
Dipsacus asper Wall.ex Henry
Triterpenoid saponins
UGT
Gene expression
