Molecular cloning, sequencing and characterization of omp48, the gene encoding for an antigenic outer membrane protein from Aeromonas veronii

• Published in: Journal of applied microbiology 2003-01, Vol.94 (5), p.908-918
• Main Authors: Vázquez‐Juárez, R.C., Barrera‐Saldaña, H.A., Hernández‐Saavedra, N.Y., Gómez‐Chiarri, M., Ascencio, F.
• Format: Article
• Language: English
• Subjects: adhesin; Aeromonas - genetics; Aeromonas - immunology; Aeromonas veronii; Amino Acid Sequence; Antigens, Bacterial - genetics; bacterial outer membrane proteins; Bacterial Outer Membrane Proteins - genetics; Bacterial Outer Membrane Proteins - immunology; Bacteriology; Base Sequence; Biological and medical sciences; cloning; Cloning, Molecular; DNA, Bacterial - genetics; Freshwater; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; Genetics; Genomic Library; inverse PCR; LamB; Microbiology; Molecular Sequence Data; Polymerase Chain Reaction - methods; Sequence Alignment
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1046/j.1365-2672.2003.01928.x

Aims: To clone, sequence and characterize the gene encoding the Omp48, a major outer membrane protein from Aeromonas veronii. Methods and Results: A genomic library of Aer. veronii was constructed and screened to detect omp48 gene sequences, but no positive clones were identified, even under low stringency conditions. The cloned gene probably was toxic to the host Escherichia coli strain, so the cloning of omp48 was achieved by inverse PCR. The nucleotide sequence of omp48 consisted of an open reading frame of 1278 base pairs. The predicted primary protein is composed of 426 amino acids, with a 25‐amino‐acid signal peptide and common Ala‐X‐Ala cleavage site. The mature protein is composed of 401 amino acids with a molecular mass of 44 256 Da. Conclusions: The omp48 gene from Aer. veronii was cloned, sequenced and characterized in detail. BLAST analysis of Omp48 protein showed sequence similarity (over 50%) to the LamB porin family from other pathogenic Gram‐negative bacteria. Significance and Impact of the Study: Bacterial diseases are a major economic problem for the fish farming industry. Outer membrane proteins are potentially important vaccine components. The characterization of omp48 gene will allow further investigation of the potential of Omp48 as recombinant or DNA vaccine component to prevent Aer. veronii and related species infections in reared fish.