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目的利用生物信息学技术对EgTK基因编码蛋白的结构和功能进行预测和分析。方法利用ExPASy系统的ProtParam、ProtScale、SignalP、SOMPA程序与SubLoc v1.0、DNAStar、IEDB、SYPEITHI、Phyre2等生物信息学软件分析EgTK的理化性质、抗原表位、跨膜区、二、三级结构等。结果 EgTK基因由7个外显子,6个内含子构成,CDS长度为1 878bp,编码625个氨基酸,分子式为C_(3015)H_(4791)N_(829)O_(900)S_(22),分子质量为67.76ku;为跨膜蛋白,位于细胞质,二级结构主要以α螺旋、无规则卷曲为主。EgTK有16个潜在的B细胞线性表位,11个CTL细胞表位,13个Th细胞表位;EgTK三级结构与人类转酮醇酶相似性为97%;经多重比对,除去两端多余序列,共有473个氨基酸残基参与比对,发现195个变异位点、123简约性信息位点、72单态突变位点。结论生物信息学技术分析EgTK蛋白存在多个B、T细胞表位,具有良好的免疫原性,可为该蛋白的基因克隆、表达等提供理论依据。
Objective To predict and analyze the structure and function of EgTK gene by bioinformatics techniques. Methods The physical and chemical properties, antigenic epitopes, transmembrane domains, secondary and tertiary structures of EgTK were analyzed by ProtParam, ProtScale, SignalP and SOMPA programs in ExPASy system and BioLab software such as SubLoc v1.0, DNAStar, IEDB, SYPEITHI and Phyre2 Wait. Results The EgTK gene consisted of 7 exons and 6 introns with a CDS of 1 878bp encoding 625 amino acids with the molecular formula C 3015 H 4791 N 829 O 900 S 22. , The molecular mass of 67.76ku; transmembrane protein, located in the cytoplasm, secondary structure mainly α-helix, random curl-based. EgTK has 16 potential B cell linear epitopes, 11 CTL cell epitopes and 13 Th cell epitopes. The EgTK tertiary structure has 97% similarity to human transketolase. After multiple alignments, both ends A total of 473 amino acid residues were involved in the alignment. 195 mutation sites, 123 parsimony informative sites, and 72 single-site mutation sites were found. Conclusions Bioinformatics analysis of EgTK protein has multiple B and T cell epitopes and has good immunogenicity, which may provide a theoretical basis for gene cloning and expression of EgTK protein.