[1]王建丽,曲洪生,尤佳,等.羊草LcCOMT基因的克隆及表达分析[J].黑龙江农业科学,2021,(12):15-23.[doi:DOI:10.11942/j.issn1002-2767.2021.12.0015]
　WANG Jian-li,QU Hong-sheng,YOU Jia,et al.Cloning and Prokaryotic Expression Analysis of LcCOMT from Leymus chinensis[J].HEILONGJIANG AGRICULTURAL SCIENCES,2021,(12):15-23.[doi:DOI:10.11942/j.issn1002-2767.2021.12.0015]
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羊草LcCOMT基因的克隆及表达分析

《黑龙江农业科学》[ISSN:1002-2767/CN:23-1204/S] 卷: 期数: 2021年12 页码: 15-23 栏目: 出版日期: 2021-12-10
Title:
Cloning and Prokaryotic Expression Analysis of LcCOMT from Leymus chinensis
作者:
王建丽1; 曲洪生2; 尤佳1; 钟鹏3; 邸桂俐1; 韩微波1; 申忠宝1
1.黑龙江省农业科学院 草业研究所，黑龙江 哈尔滨 150086；2.中商艾享生态科技股份有限公司，黑龙江 哈尔滨 150070；3.黑龙江省农业科学院 农村能源与环保研究所，黑龙江 哈尔滨 150086
Author(s):
WANG Jian-li1; QU Hong-sheng2; YOU Jia1; ZHONG Peng3; DI Gui-li1; HAN Wei-bo1; SHEN Zhong-bao1
1.Pratacultural Science Institute， Heilongjiang Academy of Agricultural Sciences， Harbin 150086，China；2.China Commerce Ax Ecological Technology Limited Company， Harbin 150070，China；3.Rural Energy and Environmental Protection Research Institute， Heilongjiang Academy of Agricultural Sciences， Harbin 150086，China
关键词:
羊草; LcCOMT; 生物信息学分析; 荧光定量表达
Keywords:
Leymus chinensis; LcCOMT; bioinformatics analysis; real-time quantitative expression
DOI:
DOI:10.11942/j.issn1002-2767.2021.12.0015
文献标志码:
A
摘要:
为初步解析羊草［Leymus chinensis(Trin.) Tzvel.］木质素合成的分子机制，依据前期转录组数据，以羊草茎cDNA为模板，克隆获得了羊草COMT基因，将其命名为LcCOMT。LcCOMT基因的开放阅读框（ORF）全长为1 065 bp，编码354个氨基酸，分子量为38.068 kDa，等电点为5.62。生物信息学分析显示，LcCOMT基因编码氨基酸序列中包含2个保守的结构域，分别为二聚化结构域与和甲基转移酶结构域，但无信号肽和跨膜结构域。系统进化分析表明，LcCOMT与大麦（Hordeum vulgare）、粗山羊草（Aegilops tauschii）的COMT基因亲缘关系最近，氨基酸序列一致性均在95％以上。同源建模发现，LcCOMT的3D模型与多年生黑麦草（Lolium perenne）LpOMT1蛋白序列相似性达到了87.85%，同源性较高，推测其可能与LpOMT1具有相似的功能。组织表达分析显示，LcCOMT在叶片、叶鞘和茎节中均有表达，但茎中表达量最高，且在同一茎节中随着生殖期的延长表达量增加。LcCOMT基因的克隆和表达分析为下一步基因功能研究奠定了基础。
Abstract:
To understand the molecular mechanisms of lignin synthesis of Leymus chinensis，LcCOMT was cloned according to RNA-sequencing data obtained in this research.The LcCOMT gene contained a 1 065 bp open reading frame(ORF) which encoded 354 amino acids，with the molecular weight of 38.068 kDa and the isoelectric point of 5.62.Bioinformatics analysis showed that LcCOMT contained two conserved domains of dimerization domain and O-Methyltransferase domain，but no signal peptide and transmembrane domain.Phylogenetic tree analysis showed that LcCOMT was closely related to the species of Hordeum vulgareand Aegilops tauschii.At the nucleotides and amino acids levels the similarity of the LcCOMT gene to the corresponding sequence in Hordeum vulgareand Aegilops tauschii was more than 95%.Homology modeling showed that the 3D model of LcCOMT was similar with that of Perennial ryegrass LpOMT1，and sequence similarity reached 87.85%，suggesting that it might have similar function to LpOMT1.Tissue specific analysis showed that LcCOMTwas expressed in leaves，leaf sheaths，and stem nodes，and with the highest expression in stems.Moreover，the expression of the same stem segment was enhanced along with the elongation.The cloning of LcCOMTand the tissue specific analysis had laid the foundation for further study of the function of this gene.

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备注/Memo

收稿日期：2021-09-13基金项目：黑龙江省省院合作项目（YS20B08）；黑龙江省科研院所科研业务费项目（CZKYF2020C007）。第一作者：王建丽（1977-），女，博士，副研究员，从事牧草分子设计与遗传育种研究。E-mail：wangjianlivip@126.com。通信作者：申忠宝（1973-），男，硕士，研究员，从事牧草分子设计与遗传育种研究。E-mail：shzhb1973@126.com。

更新日期/Last Update: 2022-02-28