Characterization of recombinant protein of Pasteurella multocida serotype B

摘要

Haemorrhagic septicaemia (HS) is an acute, fatal septicaemic bacterial disease, mainly in South and Southeast Asia, Africa and India which caused by Pasteurella multocida, a gram negative coccobacilli bacterium. The effectiveness and safety of available treatment and vaccines are limited due to antimicrobial resistance. The objective of this study was to characterize recombinant protein of Pasteurella multocida in developing vaccine against HS. Results: Bacterial culture strain was cultured in brain heart infusion (BHI) medium. Soluble proteins were extracted and separated electrophoretically using 12% gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Immunogenic soluble proteins were detected by western blotting using anti-Pasteurella serum raised in rabbit against whole cell antigens of Pasteurella multocida and anti-rabbit antibody and identified. Recombinant protein was then expressed and transformed in E.coli. Expression and purification of recombinant protein was analyzed using SDS-Page. The separation of soluble proteins showed various molecular weights on the gel, ranging from 10kDA to 170kDA. According to the western blot analysis, the most intense band detected was of approximately 28kDA and identified as lipoprotein B. Immunogenicity study of soluble protein will be carried out in response to immunogenic roles against HS and potential role in vaccine development. Purified recombinant protein size was obtained approximately 39kDa after electropherically separated using 12% SDS-Page. Conclusion: According to the immunoblotting analysis, intensity band of purified recombinant protein treated with antisera from immunized rabbit detected on film was 5398.78 with p-value equal to 0.0006 (p-value P.multocida was characterized and indicated. Further study on the immunogenicity study of recombinant protein will be carried out in response to immunogenic roles against HS and potential role in vaccine development.