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Cloning, expression, and genus-specificity analysis of 28-kDa OmpK from Vibrio alginolyticus.
- Source :
-
Journal of food science [J Food Sci] 2010 May; Vol. 75 (4), pp. M198-203. - Publication Year :
- 2010
-
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.
- Subjects :
- Amino Acid Sequence
Bacterial Outer Membrane Proteins chemistry
Bacterial Outer Membrane Proteins immunology
Bacterial Vaccines
Base Sequence
Cross Reactions
Genes, Bacterial
Molecular Sequence Data
Molecular Weight
Phylogeny
Protein Sorting Signals
Recombinant Fusion Proteins biosynthesis
Recombinant Fusion Proteins immunology
Recombinant Fusion Proteins isolation & purification
Sequence Alignment
Sequence Analysis, DNA
Species Specificity
Vibrio genetics
Vibrio immunology
Vibrio isolation & purification
Vibrio metabolism
Vibrio Infections diagnosis
Vibrio Infections prevention & control
Vibrio alginolyticus immunology
Vibrio alginolyticus isolation & purification
Bacterial Outer Membrane Proteins genetics
Bacterial Outer Membrane Proteins metabolism
Cloning, Molecular
Gene Expression
Vibrio alginolyticus genetics
Vibrio alginolyticus metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1750-3841
- Volume :
- 75
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Journal of food science
- Publication Type :
- Academic Journal
- Accession number :
- 20546410
- Full Text :
- https://doi.org/10.1111/j.1750-3841.2010.01565.x