| 苦荞FtC4H基因克隆与生物信息学分析 |
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| 引用本文： | 尹桂芳,段迎,杨晓琳,蔡苏云,王艳青,卢文洁,孙道旺,贺润丽,王莉花.苦荞FtC4H基因克隆与生物信息学分析[J].作物杂志,2022,38(1):77-197. |
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| 作者姓名： | 尹桂芳 段迎 杨晓琳 蔡苏云 王艳青 卢文洁 孙道旺 贺润丽 王莉花 |
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| 作者单位： | 1云南省农业科学院生物技术与种质资源研究所/云南省农业生物技术重点实验室/农业农村部西南作物基因资源与种质创制重点实验室,650205,云南昆明2山西中医药大学中药与食品工程学院,030619,山西太原 |
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| 基金项目： | 国家自然科学基金地区科学基金(31460379);财政部和农业农村部:国家现代农业产业技术体系(CARS-07-C-2);山西省重点研发计划项目(201803D221012-6);2019年中医药公共卫生服务补助专项“全国中药资源普查项目(财社[2019]68号)”。 |
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| 摘 要： | 克隆苦荞苯丙烷类物质代谢途径中的关键酶肉桂酸-4-羟基化酶基因（FtC4H）,为进一步研究其功能奠定基础。以云荞1号和小米荞为材料,提取不同发育期果壳RNA,利用RT-PCR法克隆苦荞FtC4H基因,运用生物信息学分析FtC4H蛋白的特征,构建FtC4H蛋白系统进化树,分析其基因表达。结果表明,克隆的FtC4H基因序列包含1299bp完整的cDNA开放阅读框,编码432个氨基酸,为亲水性不稳定碱性蛋白,具有P450超家族保守域,不具有跨膜结构域,有丰富的二级结构,三级结构预测显示FtC4H与6vby.1.A的序列相似度高。系统进化分析结果表明,本研究克隆的FtC4H与已报道的苦荞其他C4H基因不同。qRT-PCR结果表明,FtC4H在小米荞的花和叶中相对表达量显著高于云荞1号。
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| 关 键 词： | 苦荞 RT-PCR克隆 肉桂酸-4-羟基化酶 生物信息学分析 实时荧光定量PCR |
| 收稿时间： | 2021-01-18 |
Cloning and Bioinformatics Analysis of FtC4H Gene from Tartary Buckwheat |
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| Yin Guifang,Duan Ying,Yang Xiaolin,Cai Suyun,Wang Yanqing,Lu Wenjie,Sun Daowang,He Runli,Wang Lihua.Cloning and Bioinformatics Analysis of FtC4H Gene from Tartary Buckwheat[J].Crops,2022,38(1):77-197. |
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| Authors: | Yin Guifang Duan Ying Yang Xiaolin Cai Suyun Wang Yanqing Lu Wenjie Sun Daowang He Runli Wang Lihua |
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| Institution: | 1Biotechnology and Germplasm Resources Institute, Yunnan Academy of Agricultural Sciences/ Yunnan Provincial Key Laboratory of Agricultural Biotechnology/Key Laboratory of Southwestern Crop Gene Resources and Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Kunming 650205, Yunnan, China2College of Traditional Chinese Medicine and Food Engineering, Shanxi University of Chinese Medicine, Taiyuan 030619, Shanxi, China |
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| Abstract: | In this paper, the key enzyme gene cinnamic acid-4-hydroxylase (FtC4H) in the benzene propanes metabolic pathway of tartary buckwheat was cloned, laying a foundation for further research on its function. The RNA of Yunqiao 1 and Xiaomiqiao husks at different developmental stages was extracted, and the FtC4H gene from tartary buckwheat was cloned by RT-PCR, and the characteristics of FtC4H protein were analyzed by bioinformatics. The phylogenetic tree of FtC4H protein was constructed and analyzed FtC4H gene expression. The results showed that the cloned gene sequence had complete cDNA open reading frame of 1299bp, encoding 432 amino acids. FtC4H was a hydrophilic and unstable basic protein with P450 superfamily conserved domain and without transmembrane domain; The secondary structure of FtC4H was complex; the prediction of tertiary structure showed a high similarity with 6vby.1.A sequence. The results of phylogenetic analysis showed that the FtC4H cloned in this study was different from other C4H genes of tartary buckwheat reported. qRT-PCR showed that the relative expression level of FtC4H in the flowers and leaves of Xiaomiqiao were significant higher than that of Yunqiao 1. |
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| Keywords: | Tartary buckwheat RT-PCR cloning Cinnamic acid-4-hydroxylase Bioinformatics analysis qRT-PCR |
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