博落回去甲乌药碱合成酶基因的克隆及表达分析  

Cloning and Expression Analysis of Norcoclaurine Synthase(NCS) Gene in Macleaya cordata

作　　者：黄鹏[1,2] 刘金凤[1,2] 卿志星[3] 柳亦松[2,4] 唐其[1,2] 曾建国[1,2,4] Huang Peng Liu Jinfeng Qing Zhixing Liu Yisong Tang Qi Zeng Jianguo(Horticulture ＆ Landscape College, Hunan Agricultural University, Changsha, 410128 National and Local Union Engineering Research Center for the Veterinary Herbal Medicine Resources and Initiative, Hunan Agricultural University, Changsha, 410128 Veterinary Faculty of Hunan Agricultur- al University, Changsha, 410128 School of Pharmacy, Hunan University of Chinese Medicine, Changsha, 410208)

机构地区：[1]湖南农业大学园艺园林学院,长沙410128 [2]湖南农业大学兽用中药资源与中兽药创制国家地方联合工程研究中心,长沙410128 [3]湖南中医药大学药学院,长沙410208 [4]湖南农业大学动物医学院,长沙410128

出　　处：《分子植物育种》2017年第2期454-459,共6页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31200615);国家重点实验室培育基地开放课题(15KFXM09);湖南省科技厅重点项目(2014SK2005);湖南农业大学人才引进基金(13YJ09)共同资助

摘　　要：苄基异喹啉类生物碱(BIAs)是一类具有重要研究及药用价值的次生代谢产物,目前该类生物碱主要来自于植物。去甲乌药碱合成酶(NCS)参与BIAs的合成过程并发挥重要作用。为研究该基因在博落回BIAs合成过程中的作用,通过将其他物种的NCS基因序列在博落回转录组数据库中进行比对筛选到1个博落回NCS候选基因并对该序列进行生物信息学分析。经过克隆该序列中ORF区并用荧光定量PCR发现该基因的组织特异性表达规律与BIAs代谢含量相符。该研究对下一步分析该基因功能、阐明博落回NCS基因的合成机制和分子育种以及代谢工程开发新药源等方面具有重要意义。Benzylisoquinoline alkaloids（BIAs） are a kind of secondary metabolites which have important research and pharmaceutical value. Norcoclaurine synthase（NCS） plays an important role in the synthetic process of many BIAs. To investigate the role of this gene participates in synthesis of BIAs, we download the NCS gene from other species and compared the sequence with M. cordata transcriptome database and screened a candidate NCS gene from M. cordata. Then we used bioinformatics tools to analyze the sequence. We cloned the full length of its ORF region. Furthermore, we found that the tissue specificity of the gene expression regulation accorded with the metabolic levels of BIAs through the test of fluorescence quantitative PCR. The study has great significance in further analysis of the gene functions, illustration of the synthesis mechanism of M. cordata., molecular breeding and development of new medicine source in metabolic engineering.

关 键 词：博落回 去甲乌药碱合成酶 实时定量表达 

分 类 号：S567.239[农业科学—中草药栽培] Q943.2[农业科学—作物学]