Cloning, expression, and genus-specificity analysis of 28-kDa OmpK from Vibrio alginolyticus.
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.
Li
Yan-Dong
YD
Key Laboratory for Zoonosis Research, Ministry of Education, Inst. of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin Univ., Changchun 130062, PR China.
Ren
Hong-Lin
HL
Lu
Shi-Ying
SY
Zhou
Yu
Y
Han
Xue
X
Gong
Bin-Bin
BB
Zhang
Yuan-Yuan
YY
Liu
Zeng-Shan
ZS
D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D001428:Bacterial Vaccines; D001483:Base Sequence; D003001:Cloning, Molecular; D003429:Cross Reactions; D015870:Gene Expression; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D014733:Vibrio / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D014735:Vibrio Infections / Q000175:diagnosis / Q000517:prevention & control; D044149:Vibrio alginolyticus / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*
D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; C021076:protein K