论文部分内容阅读
为研究维氏气单胞菌(Aeromonasveronii)胞外产物的合成及其功能,本试验采用超滤浓缩法提取Aeromonasveronii TH0426的胞外产物,通过平板琼脂扩散法对其主要酶成分进行初步研究,对蛋白酶、脂酶、卵磷脂酶、淀粉酶、脲酶、明胶酶和溶血活性进行测定,对斑马鱼腹腔注射胞外产物测定其致病性,并对其免疫原性进行初步分析。结果显示,菌株胞外产物具有脂酶、淀粉酶、蛋白酶和溶血活性,人工感染试验结果表明,菌株胞外产物对斑马鱼具有致病性,其半数致死量为3.355 0μg/条。SDS-PAGE表明,ECP由19条蛋白带组成,利用鲤鱼抗ECP血清进行Western blot显示,组成ECP的19条蛋白带中有5条具有免疫原性,能够引发机体的免疫应答反应产生抗体,其相对分子质量为别为110 000,47 000,45 000,42 000和38 000,为Aeromonasveronii胞外产物亚单位疫苗的研制与开发提供理论依据。
In order to study the synthesis and function of extracellular products of Aeromonas veronii, the extracellular products of Aeromonas veronii TH0426 were extracted by ultrafiltration concentration, and the main enzyme components of Aeromonas veronii TH0426 were preliminarily studied by plate agar diffusion method. Protease, lipase, lecithin, amylase, urease, gelatinase and hemolytic activity were determined. The pathogenicity of zebrafish was determined by intraperitoneal injection of the extracellular product, and its immunogenicity was analyzed. The results showed that the extracellular products of the strain had lipase, amylase, protease and hemolysis activity. The artificial infection test showed that the extracellular product of the strain was pathogenic to zebrafish with the median lethal dose of 3.355 0μg / bar. SDS-PAGE showed that ECP consisted of 19 protein bands. Western blot analysis using carp anti-ECP serum showed that 5 out of 19 protein bands constituting ECP were immunogenic and could trigger the body’s immune response to produce antibodies The relative molecular mass of 110 000,47 000,45 000,42 000 and 38 000, respectively, for the Aeromonasveronii extracellular product subunit vaccine research and development to provide a theoretical basis.