药学学报, 2021, 56(2): 610-617
引用本文:
魏小艳, 李勇, 郭娟, 王雅南, 黄璐琦. 薏苡3-酮酯酰-CoA合酶基因克隆和生物信息学分析[J]. 药学学报, 2021, 56(2): 610-617.
WEI Xiao-yan, LI Yong, GUO Juan, WANG Ya-nan, HUANG Lu-qi. Cloning and bioinformatic analysis of the 3-ketoacyl-CoA synthase gene in Coix lacryma-jobi L. [J]. Acta Pharmaceutica Sinica, 2021, 56(2): 610-617.

薏苡3-酮酯酰-CoA合酶基因克隆和生物信息学分析
魏小艳1,2, 李勇2, 郭娟2, 王雅南2, 黄璐琦1,2
1. 江西中医药大学院士工作站, 江西 南昌 330004;
2. 中国中医科学院中药资源中心, 道地药材国家重点实验室培育基地, 北京 100700
摘要:
3-酮酯酰-CoA合酶(3-ketoacyl-CoA synthase,KCS)基因是调控长链脂肪酸生物合成过程中的关键基因,在薏苡的生长发育过程中起着重要的调控作用。本文以薏苡(Coix lacryma-jobi L.)作为实验材料,根据转录组测序数据,从薏苡的cDNA中克隆出KCS基因,并对其进行生物信息学分析。结果表明,薏苡KCS基因的cDNA全长序列为1 548 bp,编码515个氨基酸,生物信息学预测结果显示该基因编码蛋白质为碱性亲水不稳定蛋白,分子质量为58 608.12 Da,等电点为9.20,含有2个跨膜螺旋结构域,不含信号肽剪切位点,亚细胞定位主要在质体膜。多序列比对和进化树分析结果显示薏苡KCS与玉米、水稻、节节麦、高粱和小米等单子叶植物的同种基因具有3个相同的保守位点,而且亲缘关系更近,由此证明该基因在这些同科植物间的进化是保守的。此外,基因的表达分析结果显示KCS基因在不同油脂含量的薏苡资源中有明显的差异变化。本实验对KCS基因的克隆及其结构、性质等的研究有利于深入研究该蛋白在脂肪酸合成过程中的调控机制。
关键词:    薏苡      3-酮酯酰-CoA合酶      基因克隆      生物信息学分析     
Cloning and bioinformatic analysis of the 3-ketoacyl-CoA synthase gene in Coix lacryma-jobi L.
WEI Xiao-yan1,2, LI Yong2, GUO Juan2, WANG Ya-nan2, HUANG Lu-qi1,2
1. Academician Workstation, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China;
2. State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
Abstract:
As a key gene in the regulation of long-chain fatty acid biosynthesis, 3-ketoacyl-CoA synthase (KCS) plays an important role in the growth and development of Coix lacryma-jobi L. In this study, the KCS gene was cloned from cDNA of Coix lachryma-Jobi L. and bioinformatics analysis was performed. Results showed that the full length KCS gene was 1 548 bp encoding 515 amino acids. Bioinformatics analysis indicated that the gene encoded a 58 608.12 Da protein with an isoelectric point of 9.20 containing two transmembrane helical structure domains and lacking a signal peptide, with a likely subcellular localization in main plastid membranes. The results of multiple sequence comparisons and evolutionary tree analysis revealed that KCS had three identical conserved sequences and was closely related to KCS from monocotyledons such as Sorghum bicolor, Zea mays, Setaria italica, Panicum miliaceum, Oryza brachyantha, Hordeum vulgare, Aegilops tauschii subsp. Tauschii. We speculated that the evolution of the gene was similar among these plants of the same family. In addition, gene expression analysis showed that the KCS gene was significantly different in Coix lacryma-jobi L. isolates having different lipid content. This work will facilitate further study of the regulatory mechanism of this enzyme in fatty acid synthesis.
Key words:    Coix lacryma-jobi L.    3-ketoacyl-CoA synthase    gene cloning    bioinformatics analysis   
收稿日期: 2020-08-26
DOI: 10.16438/j.0513-4870.2020-1407
基金项目: 国家自然科学基金青年基金项目(81703657);中央本级重大增减支项目(ZZXT201702,ZZ13-YQ-085,20160302,2060302).
通讯作者: 王雅南,Tel:86-10-64087469,E-mail:wyn19870127@126.com;黄璐琦,Tel:86-10-84044340,E-mail:huangluqi01@126.com
Email: wyn19870127@126.com;huangluqi01@126.com
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