| 引用本文:任宏伟1，孟寒寒1,2，徐瑶1，赵永丽1，高婷1.药用植物珊瑚菜GlAT基因的克隆和生物信息学分析[J].世界中医药,2020,(05):. |
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| 药用植物珊瑚菜GlAT基因的克隆和生物信息学分析 |
| Cloning and Bioinformatics Analysis of Acyltransferase Gene in Medicinal Plant Glehnia littoralis（GlAT） |
| 投稿时间：2020-02-10 |
| DOI：10.3969/j.issn.1673-7202.2020.05.005 |
| 中文关键词: 药用植物 珊瑚菜 转录组 酰基转移酶 基因筛选 克隆 生物信息学分析 次生代谢 |
| English Keywords:Medicinal plant Glehnia littoralis Transcriptome Acyltransferase Gene screening Clone Bioinformatics analysis Secondary metabolism |
| 基金项目:国家自然科学基金青年项目(81903748)；上海市资源植物功能基因组学重点实验室开放基金项目（PFGR201703） |
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| 中文摘要: |
| 目的：对药用植物珊瑚菜Glehnia littoralis BAHD酰基转移酶（Acyltransferase，AT）基因序列进行生物信息学分析。方法：用茉莉酸甲酯（MeJA）处理珊瑚菜幼苗，用实时荧光定量PCR(RT-qPCR)方法检测GlAT候选基因的表达量，在已建立的珊瑚菜转录组数据中筛选出表达量较高且MeJA处理后上调表达的1条GlAT基因序列，对该基因cDNA序列进行全长克隆；根据所得的氨基酸序列进行蛋白质理化性质、二级结构、三级结构分析，利用MEGA 6.0构建该蛋白序列的NJ系统进化树。结果：GlAT的全长开放阅读框ORF（Open Reading Frame）为1 443 bp，编码480个氨基酸。该蛋白相对分子质量为52 844.14，等电点为6.92，为亲水性蛋白，属于转移酶超家族。珊瑚菜GlAT蛋白与同科植物胡萝卜Daucus carota subsp.sativus的AT蛋白亲缘关系最近。结论：首次获得珊瑚菜GlAT基因的ORF全长序列并进行了生物信息学分析，为进一步开展该基因的功能和遗传调控机制研究奠定了基础。 |
| English Summary: |
| To analyze the BAHD acyltransferase (AT) gene sequence of medicinal plants, Glehnia littoralis from bioinformatics perspective.Methods:Glehnia littoralis seedlings was treated with methyl jasmonate (MeJA).The real-time fluorescent quantitative PCR (RT-qPCR) method was used to detect the gene expression.A GlAT gene sequence with high expression and up-regulated expression after MeJA treatment were screened from established transcriptome data of Glehnia littoralis.The cDNA sequence of the gene was cloned in full-length; the physical and chemical properties, secondary structure and tertiary structure of the protein were analyzed based on obtained amino acid sequence; and NJ phylogenetic tree based on AT proteins sequence was build using MEGA 6.0 software.Results:The full-length ORF of GlAT was 1 443 bp, encoding 480 amino acid residues.The relative molecular weight of the protein was 52 844.14, and the isoelectric point was 6.92.It was a hydrophilic protein and belonged to the transferase superfamily.GlAT protein of Glehnia littoralis was closely related to the AT protein of Daucus carota subsp.sativus.Conclusion:For the first time, the full-length ORF sequence of GlAT gene from G.littoralis was obtained and bioinformatics analysis was performed, which laid foundation for further research on the function and genetic regulatory mechanism of this gene. |
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