小报春香叶醇-10羟化酶(G10H)基因克隆与生物信息学分析
首发时间:2016-11-23
摘要:香叶醇-10羟化酶(geraniol 10-hydroxylase, G10H)在植物合成萜类吲哚生物碱(monoterpenoid indole alkaloids, TIAs)的过程中充当限速酶。本研究立足于实验室所建立的小报春高通量转录组测序数据,根据从中获取的Unigene设计G10H基因引物,克隆出基因全长共1592bp,开放阅读框1500bp,编码499个氨基酸。将该氨基酸序列与其他物种G10H进行比对,构建进化树,分析其物种间进化关系,进行蛋白质生物信息学分析,包括蛋白质的结构特点、理化性质、组成及稳定性分析;跨膜区分析;信号肽识别位点;磷酸化位点;二级机构及功能;结构域分析等,这些分析为后续研究小报春G10H基因提供了理论基础。
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Cloning and Sequence Analysis of G10H Gene from Primula forbesii
Abstract:Geraniol 10-hydroxylase (G10H) is a rate-limiting enzyme in the biosynthesis of terpenoid indole alkaloids (TIAs) in plants. The database about the flowers transcriptomes of Primula forbesii were sequenced with platform of Illumina HiSeqTM 2000. The study depends on the database established by our lab. In this paper, the full length G10H of P. forbesii was cloned according to the Unigene. The gene included 1592 bp nucleotide, and the ORF (open reading frame) 1500bp, encoding 499 amino acids sequence, which was blasted with that of other plants, the phylogenetic tree was mapped and analyzed. Then bio-informatics of G10H were studied, including structure, physicochemical properties, amino acid, trans-membrane domain, signal peptide recognition, phosphorylation sites, function of secondary structure and domain analysis. Those analysis lay the foundation for the further study of G10H in P. forbesii.
Keywords: Primula forbesii G10H Bioinformatics Landscape plant
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