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为研究维氏气单胞菌(Aeromonas veronii)主要黏附素Aha1基因的生物信息学特性,本试验克隆了A.veronii TH0426主要黏附素Aha1基因片段,构建了Aha1基因序列进化树,通过生物信息学软件分析其编码蛋白的理化性质、疏水性、信号肽、跨膜区、二级结构及三级结构,并进行了Aha1蛋白的原核表达及免疫原性的检测。结果显示:Aha1基因全长1 038bp,编码345个氨基酸。TH0426株Aha1与A.veronii属同一分支,具有信号肽,跨膜区,二级结构以β折叠、无规则卷曲为主。SDS-PAGE及Western blot分析结果显示,Aha1蛋白在大肠杆菌中成功表达且其能被鼠抗黏附素阳性血清识别,表明Aha1蛋白具有一定的反应原性。以上结果为进一步研究Aha1蛋白的结构功能Aha1基因工程亚单位疫苗的制备奠定了基础。
In order to study the bioinformatics characteristics of the major adhesin Aha1 gene of Aeromonas veronii, Aha1 gene fragment of A.veronii TH0426 was cloned and the Aha1 gene sequence was cloned. Bioinformatics The physical and chemical properties, hydrophobicity, signal peptide, transmembrane region, secondary structure and tertiary structure of its encoded protein were analyzed by software, and the prokaryotic expression and immunogenicity of Aha1 protein were detected. The results showed that the Aha1 gene was 1 038 bp in length and encoded 345 amino acids. TH0426 strain Aha1 and A.veronii belong to the same branch, with signal peptide, transmembrane region, secondary structure to β-fold, random curl-based. SDS-PAGE and Western blot analysis showed that Aha1 protein was successfully expressed in Escherichia coli and recognized by mouse anti-adhesin-positive serum, indicating that Aha1 protein has a certain degree of reactogenicity. The above results laid the foundation for the further study on the construction of Aha1 gene engineering subunit vaccine with structural function of Aha1 protein.