东北雷公藤DXR HMGR基因克隆及生物信息学分析  

Cloning and Bioinformatic Analysis of DXR and HMGR Genes from Tripterygium regelii

作　　者：郭思远[1] 闫琦 李佳[1] GUO Si-yuan;YAN Qi;LI Jia(School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学中医药学院

出　　处：《中国现代中药》2019年第11期1482-1488,共7页Modern Chinese Medicine

基　　金：北京市自然科学基金项目;北京市教育委员会科技发展计划重点项目(KZ201710025022)

摘　　要：目的:克隆东北雷公藤萜类物质生物合成关键酶基因3-羟基-3-甲基戊二酰辅酶A还原酶(3-hydroxy-3-methylglutaryl coenzyme A reductase,HMGR)基因和1-脱氧-D-木酮糖-5-磷酸还原异构酶(1-deoxy-D-xylulose-5-phosphate reductoisomerase,DXR)基因全长,并对其进行生物信息学分析。方法:根据东北雷公藤转录组数据,设计特异性引物,采用聚合酶链式反应(polymerase chain reaction,PCR)技术克隆东北雷公藤DXR、HMGR基因编码区的全长序列,并进行一系列生物信息学分析。结果:成功克隆东北雷公藤DXR、HMGR基因各1条,分别命名为TrDXR和TrHMGR,两者分别长1410、1770 bp,编码469个和589个氨基酸。TrDXR为亲水性蛋白,亚细胞定位在叶绿体,无跨膜结构,转运肽定位于叶绿体,含有2个结合功能域和2个活性位点基序;TrHMGR为疏水性蛋白,亚细胞定位在内质网,有跨膜结构,无转运肽,含有4个结合功能域。TrDXR和TrHMGR与不同物种同源序列的相似性高,保守性强。结论:克隆获得TrDXR、TrHMGR基因cDNA全长,并对其生物功能进行了初步预测,为基因功能鉴定及东北雷公藤萜类物质分子形成机制研究奠定基础。Objective:To clone the full-length of 3-hydroxy-3-methylglutaryl coenzyme A reductase gene and 1-deoxy-D-xylulose-5-phosphate reductoisomerase gene related to terpenoids biosynthesis in Tripterygium regelii and analyze their bioinformatic properties.Methods:Based on the transcriptome data of T.regelii,specific primers were designed.The full-length sequences of DXR and HMGR gene coding regions were cloned by polymerase chain reaction,and a series of bioinformatics analysis were carried out.Results:One DXR and one HMGR gene were cloned,named as TrDXR and TrHMGR respectively.They are 1410 bp and 1770 bp,encoding 469 and 589 amino acids.TrDXR is a hydrophilic protein with two binding motifs and two active motifs,subcellular localization in chloroplast and no transmembrane domain,signal peptides are localized in chloroplast.TrHMGR is a hydrophobic protein contain four binding motifs,subcellular localization in cytoplasm and have transmembrane domain,no signal peptides.TrDXR and TrHMGR have high similarity and conservativeness with homologous sequences of different species.Conclusion:The full-length cDNA sequences of TrDXR and TrHMGR were cloned and the bioinformatic analysis were carried out.It laid a foundation for the identification of gene function and the study of terpenoid molecular formation mechanism in T.regelii.

关 键 词：东北雷公藤 3-羟基-3-甲基戊二酰辅酶A还原酶 1-脱氧-D-木酮糖-5-磷酸还原异构酶 基因克隆 生物信息学分析 

分 类 号：R282.71[医药卫生—中药学] Q811.4[医药卫生—中医学]