洋草麦HdCAD基因的克隆与生物信息学分析

doi:10.12160/j.issn.1672-5190.2021.03.002

畜牧与饲料科学 ›› 2021, Vol. 42 ›› Issue (3): 7-13.doi: 10.12160/j.issn.1672-5190.2021.03.002

洋草麦HdCAD基因的克隆与生物信息学分析

郝丽芬, 房永雨, 孙林, 赵俊利, 史志丹, 詹鹤年, 丁海君

内蒙古自治区农牧业科学院,内蒙古呼和浩特 010031

收稿日期:2021-04-22 出版日期:2021-05-30 发布日期:2021-06-21
通讯作者: 丁海君（1979—）,女,研究员,博士,主要从事牧草种质资源与育种研究工作。
作者简介:郝丽芬（1987—）,女,助理研究员,博士,主要从事植物分子生物学研究工作。
基金资助:
内蒙古自治区农牧业创新基金项目（2020CXJJM09）; 内蒙古自治区自然基金与内蒙古农牧业科学院青年联合基金项目[2016MS（LH）0305]

Cloning and Bioinformatic Analysis of HdCAD Gene in Hordeum distichon

HAO Li-fen, FANG Yong-yu, SUN Lin, ZHAO Jun-li, SHI Zhi-dan, ZHAN He-nian, DING Hai-jun

Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences,Hohhot 010031,China

Received:2021-04-22 Online:2021-05-30 Published:2021-06-21

摘要/Abstract

摘要： [目的]克隆洋草麦肉桂醇脱氢酶基因（HdCAD）,揭示HdCAD基因的序列特性,对HdCAD蛋白进行生物信息学分析。[方法]以洋草麦转录组数据中的HdCAD为参考序列,设计1对特异引物,利用RT-PCR技术克隆HdCAD基因;利用生物信息学软件对HdCAD基因序列进行分析,对HdCAD蛋白的理化性质、蛋白结构、亚细胞定位等进行预测和系统进化树分析。[结果]HdCAD基因编码区全长1 071 bp,共编码356个氨基酸。HdCAD蛋白含有17个磷酸化位点,不含有跨膜结构域和信号肽位点,预测定位在细胞质中;二级结构中,以无规卷曲元件占比最高;含有1个典型的CAD1结构域,属于MDR超家族。系统进化树分析结果显示,HdCAD蛋白与二粒小麦、节节麦、西藏裸大麦亲缘关系最近。[结论]该试验成功克隆HdCAD基因、预测了该基因编码蛋白的序列特征和功能,可以为今后研究洋草麦抗非生物胁迫功能提供参考。

关键词: 洋草麦, 肉桂醇脱氢酶, 基因克隆, 生物信息学分析

Abstract: [Objective] To reveal the sequence characteristics of cinnamyl alcohol dehydrogenase gene （HdCAD） in Hordeum distichon. [Method] A pair of specific primers designed by referring HdCAD in transcriptome data of Hordeum distichon were used to clone HdCAD gene by RT-PCR. The gene sequence of HdCAD and HdCAD protein sequence encoded by this gene were bioinformatically analyzed. [Result] The total length of the coding region of HdCAD gene was 1 071 bp, encoding 356 amino acids in total. The HdCAD protein contained 17 phosphorylation sites, and the predicted location was in cytoplasm. There were no transmembrane domains and signal peptide sites in HdCAD protein. In the secondary structure, random coiled elements accounted for the highest proportion. The HdCAD protein contained a typical CAD1 domain which belonged to the MDR superfamily. The phylogenetic tree demonstrated that HdCAD protein was closely related to Triticum dicoccum, Aegilops tauschii and Tibet naked barley.[Conclusion] The HdCAD gene is successfully cloned, and the sequence characteristics and functions of its encoding protein are predicted by bioinformatics analysis, which provides a basis for researches on resistance to abiotic stress of Hordeum distichon.

Key words: Hordeum distichon, cinnamyl alcohol dehydrogenase, gene cloning, bioinformatics analysis

中图分类号:

郝丽芬, 房永雨, 孙林, 赵俊利, 史志丹, 詹鹤年, 丁海君. 洋草麦HdCAD基因的克隆与生物信息学分析[J]. 畜牧与饲料科学, 2021, 42(3): 7-13.

HAO Li-fen, FANG Yong-yu, SUN Lin, ZHAO Jun-li, SHI Zhi-dan, ZHAN He-nian, DING Hai-jun. Cloning and Bioinformatic Analysis of HdCAD Gene in Hordeum distichon[J]. Animal Husbandry and Feed Science, 2021, 42(3): 7-13.

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洋草麦HdCAD基因的克隆与生物信息学分析

Cloning and Bioinformatic Analysis of HdCAD Gene in Hordeum distichon

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