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[目的]:克隆鳗弧菌flaE基因并分析其蛋白结构,为研究该蛋白的生物学功能及免疫原性奠定基础。[方法]通过PCR方法对鳗弧菌flaE基因进行扩增,利用生物学软件对其序列及蛋白质结构进行分析。[结果]所得flaE基因的大小为841bp,其开放阅读框长度为798 bp,编码262个氨基酸。蛋白分子质量为28 411.4,理论等电点p I为5.70,脂肪系数为81.60,不稳定系数为31.81;为疏水性蛋白;没有信号肽和跨膜螺旋结构;保守区结构域属于flagellin家族;二级结构中以α螺旋为主,其次是无规则卷曲和β片层,少量β转角;三级结构与3k8v.1.A的结构模型相似率为90%。[结论]成功克隆了鳗弧菌flaE基因,Gen Bank登录号为KM091934。
[Objective] The research aimed to clone flaE gene of Vibrio anguillarum and analyze its protein structure, which laid the foundation for studying the biological function and immunogenicity of this protein. [Method] The flaE gene of Vibrio anguillarum was amplified by PCR and its sequence and protein structure were analyzed by using biological software. [Result] The obtained flaE gene was 841 bp in length and its open reading frame was 798 bp in length, encoding 262 amino acids. The protein molecular mass was 28 411.4, the theoretical isoelectric point p I was 5.70, the fat coefficient was 81.60, the instability coefficient was 31.81; it was a hydrophobic protein; there was no signal peptide and transmembrane helix structure; the conserved region belonged to flagellin family; The α-helix was the main structure in the class structure, followed by random curl and β-sheet with a small amount of β-angle. The similarity between the tertiary structure and the structural model of 3k8v.1.A was 90%. [Conclusion] The flaE gene of Vibrio anguillarum was successfully cloned. The Gen Bank accession number was KM091934.