| 多穗柯黄酮3-羟化酶基因的克隆与序列分析 |
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| 引用本文： | 尹峰,龙月红,冯若宣,林丽梅,黄剑,邢朝斌. 多穗柯黄酮3-羟化酶基因的克隆与序列分析[J]. 中草药, 2017, 48(24): 5085-5089 |
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| 作者姓名： | 尹峰 龙月红 冯若宣 林丽梅 黄剑 邢朝斌 |
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| 作者单位： | 华北理工大学生命科学学院, 河北 唐山 063210,华北理工大学生命科学学院, 河北 唐山 063210,华北理工大学生命科学学院, 河北 唐山 063210,华北理工大学生命科学学院, 河北 唐山 063210,百色学院农业与食品工程学院, 广西 百色 533000,华北理工大学生命科学学院, 河北 唐山 063210 |
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| 基金项目： | 国家自然科学基金项目（31760221）；华北理工大学培育基金（SP201508）；华北理工大学大学生创新创业训练计划（X2017185） |
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| 摘 要： | 目的克隆多穗柯Lithocarpus polystachyus黄酮3-羟化酶(flavanone 3-hydroxylase,F3H)基因,了解其基因特征并初步探明其在各器官中的表达情况。方法分别提取多穗柯叶片的总RNA及基因组DNA,根据转录组测序结果,设计特异性引物,PCR扩增得到多穗柯F3H基因的c DNA及DNA序列,测序后进行生物信息学分析,通过qRT-PCR法检测多穗柯F3H基因在不同器官中的表达情况。结果多穗柯F3H基因c DNA全长1 340 bp,包含长1 092 bp的开放阅读框,编码363个氨基酸的蛋白质,定位于细胞质中。qRT-PCR结果表明F3H基因在多穗柯各器官中均有表达,但表达量具有显著性差异(P0.05)。结论首次对多穗柯F3H基因进行了克隆和生物信息学分析,证实F3H基因在多穗柯不同器官中表达量差异显著,为多穗柯中黄酮类的次生代谢研究奠定了基础。
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| 关 键 词： | 多穗柯 黄酮3-羟化酶 克隆 生物信息 qRT-PCR 表达分析 |
| 收稿时间： | 2017-09-13 |
Cloning of flavanone 3-hydroxylase gene from Lithocarpus polystachyus and its sequence analysis |
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| YIN Feng,LONG Yue-hong,FENG Ruo-xuan,LIN Li-mei,HUANG Jian and XING Zhao-bin. Cloning of flavanone 3-hydroxylase gene from Lithocarpus polystachyus and its sequence analysis[J]. Chinese Traditional and Herbal Drugs, 2017, 48(24): 5085-5089 |
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| Authors: | YIN Feng LONG Yue-hong FENG Ruo-xuan LIN Li-mei HUANG Jian XING Zhao-bin |
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| Affiliation: | College of Life Sciences, North China University of Science and Technology, Tangshan 063210, China,College of Life Sciences, North China University of Science and Technology, Tangshan 063210, China,College of Life Sciences, North China University of Science and Technology, Tangshan 063210, China,College of Life Sciences, North China University of Science and Technology, Tangshan 063210, China,Agricultural and Food Engineering, Baise University, Baise 533000, China and College of Life Sciences, North China University of Science and Technology, Tangshan 063210, China |
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| Abstract: | Objective To clone flavanone 3-hydroxylase (F3H) gene from Lithocarpus polystachyus, and to understand its gene characteristics and initially investigate its expression level in different organs. Methods The total RNA and genomic DNA from blade of L. polystachyus were extracted. Based on the result of RNA-seq, a pair of specific primers were designed. cDNA and DNA sequences of F3H gene from L. polystachyus were amplified by PCR, then bioinformation analysis was performed after sequencing. The expression level of F3H gene in different organs of L. polystachyus was detected by qRT-PCR. Results The full length of cDNA of F3H gene was 1 340 bp containing a 1 092 bp open reading frame that encoded 393 amino acids, and F3H was located in the cytoplasm. The result of qRT-PCR showed that F3H gene expressed in different organs of L. polystachyus, and the expression levels of F3H gene were significantly different in different organs (P < 0.05). Conclusion The F3H gene of L. polystachyus was cloned and its bioinformation was analyzed for the first time, proving that the expression level of F3H gene in different organs of L. polystachyus was different. This finding lays a foundation for the studies on secondary metabolism of flavonoids in L. polystachyus. |
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| Keywords: | Lithocarpuspolystachyus Rehd flavanone 3-hydroxylase clone bioinformation qRT-PCR expression analysis |
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