Analysis And Application Of Outer Membrane Protein Of Pesudomonas Putida

Pseudosciaena crocea is the main farmed species of the Fujian Province,annual output of 90,000 tons,China accounted for 90%of production.In recent years,breeding Pseudosciaena crocea in the spring of low temperature(about 15℃or 13℃～18℃)is often caused by the outbreak of Pseudomonas mainly stench to the Pseudosciaena crocea breeding industry has brought huge losses.This study was extracted with non-ionic detergent method Pseudomonas putida outer membrane protein,SDS-PAGE electrophoresis showed 49 kDa,40 kDa,26 kDa protein as its main protein band;to extract the outer membrane protein antigen prepared 8C4,8H7,8F9 three monoclonal antibody titers were 1:3200.1:12800,1:800,while in the outer membrane proteins of Pseudomonas putida Pseudosciaena crocea immune serμm obtained Pseudosciaena crocea multi-resistant,the most efficient price of 1:102400;Western-blotting including these two antibodies Pseudomonas putida,including six outer membrane proteins of aquatic pathogens analysis shows that.8C4 and Plesiomonas shigelloides’OMPs has two immunity strip at 85 kDa and 62 kDa OMPs,8F9 in these two did not respond,two monoclonal antibodies,there is little difference in the antigenic site recognition;anti-mouse anti-fish representing more immunoreactive bands in Western-blotting reaction,described two animals there are some differences in terms of antigen presentation.However,these types of antibodies can recognize different pathogens,indicating that these bacteria have common epitopes,provides the basis for the study of a broad spectrum of subunit vaccines.In order to explore the immune disease prevention and control about the Pseudomonas putida,other than this study membrane protein antigen with the company SEPPIC MONTANIDE ISA 763 A VG is made vaccine adjuvant.In the field of application of the experiment,each Pseudosciaena crocea intraperitoneal injection of 100μg membrane protein vaccine after immunization 25 d,pathogens 2 × 103 cfu/dose challenge experiments fish tail,after the attack 12 d,outer membrane protein group of ISA 763 plus the survival rate was 57%,pure outer membrane protein immunized group survival rate was 26%in the control group,the survival rate was 6%,the outer membrane protein adjuvant ISA 763 small group of immune protection for the control group was 55%,indicating that the stench false Aeromonas outer membrane protein has good immunogenicity,the vaccine prepared for the Pseudosciaena crocea improved immunity to bacteria,Pseudomonas putida can be used for disease prevention and control.