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为了研究维氏气单胞菌胞外产物在青虾感染中的致病作用以及维氏气单胞菌的致病机理,本试验以青虾源维氏气单胞菌QXF0711B为研究对象,提取其胞外产物,采用打孔法测定其胞外产物的酶活性,并对溶血活性进行溶血谱分析,同时分析其致病性。应用液相质谱与串联质谱相结合方法(LC-MSMS)对其胞外产物蛋白成分进行鉴定,利用Gene Ontology(GO)对已鉴定蛋白进行生物学过程、分子功能和细胞组分的分类分析。结果表明:QXF0711B菌株胞外产物具有淀粉酶活性、脂肪酶活性、蛋白酶活性和溶血活性,不具有明胶酶活性;其可溶解多种动物红细胞,尤以对鱼类红细胞溶血性更强,但对鸡、鸭红细胞无溶血活性。通过对菌株胞外产物蛋白成分分析显示有90种蛋白:共参与45种生物学过程,主要涉及碳水化合物代谢过程、运输等;共有46种分子功能,主要涉及DNA结合、ATP结合、金属离子结合等;为12种细胞组分,主要包括膜的有机组成、细胞质、细胞外膜等。
In order to study the pathogenesis of Aeromonas sobria extracellular products in the infection of freshwater shrimp and pathogenic mechanism of Aeromonas sobria, freshwater shrimp Aeromonas sobria QXF0711B was used as the research object to extract The extracellular products, using punch method to determine the extracellular enzyme activity, and hemolytic activity hemolysin spectrum analysis, and analysis of its pathogenicity. The protein components of extracellular products were identified by liquid chromatography-tandem mass spectrometry (LC-MSMS), and the biological processes, molecular functions and cell components were identified by Gene Ontology (GO). The results showed that the extracellular products of QXF0711B strain had amylase activity, lipase activity, protease activity and hemolytic activity without gelatinase activity. It could dissolve red blood cells of a variety of animals, especially hemolytic of fish erythrocytes, Chicken, duck red blood cells without hemolytic activity. Analysis of the protein components of the extracellular products of the strains showed that 90 proteins were involved: 45 involved in a total of 45 biological processes, mainly involved in carbohydrate metabolism, transport, etc .; a total of 46 molecular functions, mainly involving DNA binding, ATP binding, metal ion binding Etc .; for 12 kinds of cellular components, including the organic membrane, cytoplasm, extracellular membranes and so on.