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目的扩增和测序肠出血型大肠埃希菌O157:H7 tir基因,利用生物信息学预测分析其结构和功能特征,以探讨Tir作为疫苗候选抗原的可能性。方法利用PCR技术扩增tir基因并测序,应用生物信息学网站在线分析工具和vector NT Isuite软件分析Tir蛋白结构和生物学功能,预测B细胞抗原表位。结果该基因全长1674bp,编码558个氨基酸,蛋白质总体亲水性高,有稳定的理化性质。含有3个结构和功能域,两段穿膜结构,多个磷酸化位点,预测20个B细胞线性表位。结论Tir毒力因子是很有前景的疫苗候选抗原,为肠出血型大肠埃希菌O157:H7的疫苗研究提供了理论依据。
OBJECTIVE: To amplify and sequence the enterohemorrhagic Escherichia coli O157: H7 tir gene by bioinformatics and to predict the structure and function of E.coli O157: H7 tir, and to explore the possibility of using Tir as vaccine candidate antigen. Methods The tir gene was amplified by PCR and sequenced. The bioinformatics website online analysis tool and vector NT Isuite software were used to analyze the Tir protein structure and biological function, and predict the B cell epitopes. Results The gene was 1674 bp in length and encoded 558 amino acids. The protein was highly hydrophilic and had stable physicochemical properties. Contains three structures and functional domains, two transmembrane structures, multiple phosphorylation sites, and predicts 20 B-cell linear epitopes. Conclusion Tir virulence factor is a promising vaccine candidate antigen, providing a theoretical basis for the vaccine research of enterohemorrhagic Escherichia coli O157: H7.