博落回中四氢小檗碱氧化酶McSTOX基因的克隆及表达分析  

Cloning and Expression Analysis of Tetrahydroberberine Oxidase(STOX) 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] 

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：《基因组学与应用生物学》2017年第6期2458-2463,共6页Genomics and Applied Biology

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：小檗碱是一种具有重要药用价值的次生代谢产物,目前该化合物主要来自于植物。四氢小檗碱氧化酶(STOX)是合成小檗碱的最后一步反应的酶,其在合成小檗碱的过程中发挥着关键的作用。本研究通过博落回转录组数据筛选到1个小檗碱氧化酶候选基因,并通过PCR技术对该序列进行克隆并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小檗碱代谢规律一致。该研究对下一步分析该基因功能、阐明博落回中STOX基因的合成机制和下一步运用植物代谢工程技术提高小檗碱在博落回中的含量具有重要意义。Berberine is a kind of secondary metabolite that has important pharmaceutical value, which is mainly derived from plants at present. Tetrahydroberberine oxidase is the final enzyme in berberine biosynthesis and plays a key role in the synthesis process of berberine. The study screened a candidate STOX gene from M. cordata transcriptome database, and we cloned this sequence using PCR techniques and analyzed it by bioinformatics tools. We cloned the full length of its ORF region and found that the tissue specific expression pattern of the gene accorded with the metabolic regulation of berberine through the test of fluorescence quantitative PCR. This study had great significance in further analysis of the gene functions, illustration of the synthesis mechanism of M. cordata and using plant metabolic engineering technology to enhance the content ofberberine in M. eordata.

关 键 词：博落回 四氢小檗碱氧化酶 实时定量表达 

分 类 号：Q943.2[生物学—植物学]