摘要
目的鉴定并克隆药用植物马缨丹体内活性成分三萜皂苷生物合成途径的关键限速酶基因2,3-环氧角鲨烯环化酶基因。方法基于序列比对法,从马缨丹的转录组数据中鉴定出候选2,3-环氧角鲨烯环化酶基因,利用生物信息学软件分析其序列特征、预测其编码蛋白的生化特征及其进化关系,设计特异性引物从马缨丹cDNA中扩增该基因并构建克隆载体。结果成功鉴定出一个2,3-环氧角鲨烯环化酶基因LaOSC1,该基因具备OSC基因特有的保守功能域。进化分析结果提示其可能编码β-AS蛋白,可催化2,3-环氧角鲨烯形成β-香树脂醇,并成功构建了LaOSC1的克隆载体。结论成功鉴定并克隆了马缨丹中一个2,3-环氧角鲨烯环化酶基因LaOSC1,为后续对马缨丹体内活性成分的生物合成途径研究奠定了基础。
Objective To identify and clone candidate 2,3-oxidosqualene cyclase gene,a rate-limiting enzyme involved in the biosynthetic pathway of active triterpenoid components,in the medicinal plant Lantana camara.Methods Candidate squalene 2,3-oxidosqualene cyclase gene was identified from transcriptome data of Lantana camara based on sequence similarity comparison.Its sequence characteristics,biological characteristics of its coding protein,and evolutionary relationship were analyzed by bioinformatics softwares.Specific primers were designed to amplify the squalene 2,3-oxidosqualene cyclase gene from cDNA of Lantana camara and construct the clone vector.Results A 2,3-oxidosqualene cyclase gene LaOSC1 with the conserved domain of OSC gene was successfully identified,whose coding protein might catalyze the formation of beta-amyrin from 2,3-oxidosqualene cyclase.The phylogenetic tree suggested that it might encodeβ-amyrin synthase protein,and the cloning vector of LaOSC1 was successfully constructed.Conclusion LaOSC1,a 2,3-oxidosqualene cyclase gene from Lantana camara,is successfully identified and cloned,which lays a foundation for the subsequent study of the biosynthetic pathway of active components in Lantana camara.
作者
高蝶娜
范志高
冯时茵
GAO Die-na;FAN Zhi-gao;FENG Shi-yin(The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405)
出处
《中南药学》
CAS
2024年第4期955-960,共6页
Central South Pharmacy
