| 摘要: |
| 为探究夏枯草中GGPPS基因的生物学特性及功能,该文在夏枯草转录组测序的基础上设计特异性引物,采用逆转录 PCR 技术获得夏枯草中GGPPS基因的全长核苷酸序列,并进行生物信息学分析; 采用 qPCR 法分析PvGGPPS基因在不同外源性物质诱导下在夏枯草果穗中的表达量以及该基因在夏枯草不同组织中的表达量。结果表明:PvGGPPS基因开放阅读框1 092 bp,编码363个氨基酸,理论分子量为38 815.68 D,等电点为5.69。PvGGPPS蛋白具有异戊烯基焦磷酸合酶家族的特征结构域。系统进化树表明PvGGPPS蛋白与丹参、毛喉鞘蕊花GGPPS蛋白具有较高的亲缘关系。qPCR分析表明,PvGGPPS基因在叶中表达量高于果穗及茎。对果穗施加7种外源性物质处理24 h后,GA3 处理组该基因表达量升高。PvGGPPS基因在夏枯草不同组织中表达量差异较大,且受外源物质诱导表达。该研究结果为进一步研究PvGGPPS基因对夏枯草萜类成分合成途径中的功能及表达调控奠定基础。 |
| 关键词: 夏枯草, PvGGPPS, 基因克隆, 诱导表达, 表达分析 |
| DOI：10.11931/guihaia.gxzw201906006 |
| 分类号:Q943 |
| 文章编号:1000-3142(2020)06-0882-09 |
| Fund project:国家自然科学基金(81603232); 国家重点研发计划项目(2017YFC1702800); 河南省重大科技专项项目(171100310500); 河南中医学院博士科研基金(BSJJ2015-13); 中央引导地方科技发展专项项目 [Supported by the National Natural Science Foundation of China(81603232); National Key Research and Development Plan of China(2017YFC1702800); Major Science and Technology Program in Henan Province(171100310500); Doctor Research Fund of Henan College of Traditional Chinese Medicine(BSJJ2015-13); Special Fund for Central Guiding Local Scientific and Technological Development]。 |
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| Cloning and induced expression analysis of PvGGPPS gene in Prunella vulgaris |
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ZHANG Mengjia1, DONG Chengming1,2, ZHU Yunhao1,2*
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1. School of Pharmacy, Henan University of Traditional Chinese Medicine, Zhengzhou 450046, China;2. Henan Province Collaborative Innovation
Center of Respiratory Disease Diagnosis and Treatment and New Drug Research and Development, Zhengzhou 450046, China
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| Abstract: |
| In order to explore the biological characteristics and functions of GGPPS gene in Prunella vulgaris, the specific primers were designed based on the sequencing of P. vulgaris transcriptome. The full-length nucleotide sequence of GGPPS gene was obtained in P. vulgaris by reverse transcription PCR technology, and the bioinformatics analysis was done. qPCR was used to analyze the expression of PvGGPPS gene in ear induced by different exogenous substances and in different organs in P. vulgaris. The results showed that the PvGGPPS gene had an open reading frame of 1 092 bp and encoded 363 amino acids, with a theoretical molecular weight of 38 815.68 D and an isoelectric point of 5.69. PvGGPPS protein had the characteristic domain of isopentenyl pyrophosphate synthase family. Phylogenetic tree showed that PvGGPPS protein was closely related to Salvia miltiorrhiza and Coleus forskohlii GGPPS protein. qPCR analysis showed that the expression of PvGGPPS gene in leaves was higher than that in ears and stems. The expression level of PvGGPPS gene was increased in the ear after GA3 treatment for 24 h, one of seven exogenous substances treated. The expression of PvGGPPS gene in different tissues of Prunella vulgaris was quite different and was induced by exogenous substances treatment. The results of this study lay a foundation for further study on the function and expression regulation of PvGGPPS gene in the synthesis pathway of terpenoids from P. vulgaris. |
| Key words: Prunella vulgaris, PvGGPPS, gene cloning, induced expression, expression analysis |
