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Cloning, expression, and genus-specificity analysis of 28-kDa OmpK from Vibrio alginolyticus
Yan-Dong Li1, Hong-Lin Ren, Shi-Ying Lu
1Key Laboratory for Zoonosis Research, Ministry of Education, Inst. of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin Univ., Changchun 130062, PR China.
Abstract:
Some Vibrio species are universal marine pathogens and Vibrio infections are often encountered due to consumption of raw or uncooked seafood. The outer membrane proteins, playing a key role in interaction between bacteria and hosts, are potential candidates for development of vaccine and markers of the genus Vibrio. In this study, the ompK (outer membrane protein K) genes of Vibrio alginolyticus, V. vulnificus, V. parahaemolyticus, V. fluvialis, and V. mimicu were cloned with 798 to 822 nucleotides encoding 266 to 274 amino acids. The ompK gene from V. alginolyticus was expressed in Escherichia coli using pET-22b expression vector. The recombinant fusion OmpK protein with 6xHis tag was purified with nickel chelate affinity chromatography. The polyclonal antibody (titer, 1:102400) against V. alginolyticus OmpK was developed in guinea pigs and it positively reacted with each of 5 Vibrio species but negative to other 18 Gram-negative bacterial strains. The result suggests that Vibrio OmpK protein could be a genus-specific antigen, which can be used for developing vaccines and rapid detection of multiple Vibrio species.
Insights
Outer membrane protein K (OmpK) from Vibrio species shows potential as a genus-specific antigen. This discovery could lead to new vaccines and rapid detection methods for Vibrio infections.
Area of Science:
- Microbiology
- Immunology
- Bacteriology
Background:
- Vibrio species are significant marine pathogens causing infections through seafood consumption.
- Outer membrane proteins are crucial for bacterial-host interactions and potential vaccine targets.
Purpose of the Study:
- To investigate the outer membrane protein K (OmpK) of five Vibrio species as a potential genus-specific antigen.
- To explore the utility of OmpK for vaccine development and rapid detection of Vibrio.
Main Methods:
- Cloning and sequencing of ompK genes from five Vibrio species.
- Expression and purification of recombinant OmpK protein from Vibrio alginolyticus in E. coli.
- Development of a polyclonal antibody against V. alginolyticus OmpK and testing its reactivity.
Main Results:
- The ompK genes varied slightly in size (798-822 nucleotides).
- A recombinant OmpK protein with a 6xHis tag was successfully purified.
- The developed polyclonal antibody showed high specificity, reacting positively with five Vibrio species and negatively with 18 other Gram-negative bacteria.
Conclusions:
- Vibrio OmpK protein is a promising genus-specific antigen.
- OmpK can be utilized for the development of broad-spectrum vaccines against Vibrio infections.
- OmpK serves as a potential marker for the rapid detection of multiple Vibrio species.
