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Published on: January 27, 2021
[Construction and characterization of ompH gene knockout mutant of avian Pasteurella multocida C48-3]
Wulumuhan Nazierbieke1, Yufeng Zhang, Fengjuan Gong
1College of Biology and Environmental Sciences, Jishou University, Jishou 416000, China. ulum@jsu.edu.cn
Objective:
To study the role of the outer membrane protein H (OmpH) in pathogenicity of avian Pasteurella multocida.
Methods:
The ompH knock-out mutant of avian P. multocida C48-3 was constructed by homologous recombination. The DNA replacement was confirmed by PCR, RT-PCR and Western blot. We compared the differences of biological characteristics such as growth rate, capsular structure, adhesion ability and virulence between the ompH knockout mutant of C48-3 Delta ompH and parent strain C48-3, as well as the complemented strain C48-3C.
Results:
C48-3 Delta ompH was successfully constructed. Electron microscopy examination of C48-3 Delta ompH shows the absence of capsular material compared to the parent strain C48-3 and complemented strain C48-3C. The adhesion assay shows that the number of C48-3 Delta ompH adhered to CEF cells was significantly lower than that of C48-3 and C48-3C. C48-3 Delta ompH was relatively attenuated in mice by intraperitoneal injection.
Conclusion:
The construction of C48-3 Delta ompH would facilitate further study on pathogenesis of avian Pasteurella multocida.
Insights
The outer membrane protein H (OmpH) is crucial for avian Pasteurella multocida pathogenicity. Deleting OmpH reduced bacterial adhesion and virulence, impacting avian disease studies.
Area of Science:
- Bacteriology
- Microbial Pathogenesis
- Avian Health
Context:
- Avian Pasteurella multocida is a significant pathogen in poultry.
- Outer membrane proteins play vital roles in bacterial virulence.
- Understanding OmpH's function is key to controlling P. multocida infections.
Purpose:
- To investigate the role of outer membrane protein H (OmpH) in the pathogenicity of avian P. multocida.
- To construct and characterize an OmpH knockout mutant of P. multocida.
Summary:
- An OmpH knockout mutant (C48-3 Delta ompH) of avian P. multocida was successfully created using homologous recombination.
- The mutant exhibited reduced capsular material, significantly lower adhesion to host cells (CEF), and attenuated virulence in a mouse model compared to the parent strain.
- These findings highlight OmpH's importance in P. multocida's biological characteristics and pathogenicity.
Impact:
- The generated OmpH knockout mutant provides a valuable tool for further research into the pathogenesis of avian P. multocida.
- This study contributes to understanding bacterial virulence factors, potentially informing strategies for disease control in poultry.
