| 引用本文:李志红1, 2，杨萌萌1, 2，徐静雅1, 2，凡贞洁1, 2，王瑶瑶1, 2，王龙1, 2，侯典云1, 2.山茱萸bHLH2基因的克隆与分析[J].世界中医药,2020,(05):. |
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| 山茱萸bHLH2基因的克隆与分析 |
| Cloning and Analysis of Transcription CobHLH2 Genes in Cornus officinalis |
| 投稿时间：2020-02-10 |
| DOI：10.3969/j.issn.1673-7202.2020.05.007 |
| 中文关键词: 山茱萸 转录组 转录因子 CobHLH2 基因 克隆 生物信息学 分析 |
| English Keywords:Cornus officinalis Transcriptome Transcription factors CobHLH2 Gene Cloning Bioinformatics Analysis |
| 基金项目:国家自然科学基金资助项目（U1404829）；西藏自治区重点科技计划项目（2015—ZD—NM—0） |
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| 中文摘要: |
| 目的：获得山茱萸转录因子CobHLH2的cDNA序列。方法：以山茱萸转录组测序数据中的c101916_g1序列为参考，利用Primer Premier 5.0软件在其开放阅读框两端设计特异性引物，把山茱萸果实总RNA反转录获得的cDNA作模板，通过PCR扩增、纯化回收和克隆测序，获得CobHLH2序列，并经生物信息学分析初步了解其基本结构与功能。结果：CobHLH2的ORF长度为795 bp，共编码264个氨基酸，为亲水性蛋白。CobHLH2蛋白的二级结构以无规卷曲和α-螺旋为主，有少量的β-转角和延伸链。多序列比对和系统进化树分析结果表明，CobHLH2的氨基酸序列与猕猴桃和茶树的bHLH类转录因子同源性较高。结论：首次获得山茱萸CobHLH2转录因子的cDNA序列，并进行了生物信息学分析，为深入研究bHLH类转录因子的结构和功能奠定基础。 |
| English Summary: |
| To obtain the cDNA sequence of CobHLH2 transcription factor in Cornus officinalis.Methods:Based on the c101916_g1 sequence from the transcriptome data of C.officinalis as the reference, a pair of specific primers were designed at the 2 ends of its open reading frame using Primer Premier 5.0 software.The total RNA of Cornus officinalis fruits was extracted and reversed transcribed into cDNA as the model.The CobHLH2 sequence was obtained by PCR amplification, purification recovery and clone sequencing.Its basic structure and function through bioinformatics analysis were understood.Results:The ORF of CobHLH2 was 795 bp in length, encoding 264 amino acids and it was a hydrophilic protein.The secondary structure of CobHLH2 protein mainly was Random curl and α-helix, and β-turns and extended chains were less.The analysis of multiple sequence alignments and phylogenetic trees showed that the amino acids sequence of CobHLH2 shared high homology with bHLH transcription factors of Camellia sinensis and Actinidia chinensis.Conclusion:The cDNA sequence of CobHLH2 from C.officinalis was cloned for the first time and bioinformatics analysis was performed, which would lay the foundation for further study on the structure and function of bHLH transcription factors. |
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