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)

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1443 bp,编码480个氨基酸。该蛋白相对分子质量为52844.14,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。

Objective: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 1443 bp,encoding 480 amino acid residues.The relative molecular weight of the protein was 52844.14,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.