ISOLATION, CLONING AND PHYLOGENETIC ANALYSIS OF pld AND cp40, VIRULENCE FACTORS OF A MEXICAN ISOLATE OF Corynebacterium pseudotuberculosis ovis

Abstract

<p><strong>Background. </strong>Caseous lymphadenitis is a worldwide distribute disease that affects the sheep and goat industry. <em>Corynebacterium pseudotuberculosis</em> <em>ovis</em> is a facultative intracellular Gram-positive bacterium, considered the etiologic agent of the disease. Complete genome sequences of Mexican isolates have been obtained and different strain has been previously characterized. The study of virulence factors allows establishing potential candidates for the development of vaccines and diagnostic tools. <strong>Objective.</strong> To identify the complete <em>pld </em>and <em>cp40</em> genes sequence from a Mexican isolate of <em>Corynebacterium pseudotuberculosis ovis</em>, principal virulence factors. <strong>Methodology</strong>. <em>Corynebacterium pseudotube</em>rculosis isolate 2J-L was obtained from an abscess of a sheep of the State of Jalisco. The complete <em>pld </em>and <em>cp40</em> genes were amplified by PCR, cloned into a replicative vector and sequenced by Sanger automatic sequencing. Gene sequences conservancy was established, analyzing homology across previously reported genes of Mexican strains MEX1, MEX2, MEX9, MEX25 and MEX29. <strong>Results</strong>. Sequences of <em>pld</em> and <em>cp40</em> genes from isolate 2J-L presented high percentages of similarity (99%) in comparison with the sequences of other isolates of <em>Corynebacterium pseudotuberculosis</em>, reported in the GenBank database. The analysis of nucleotide sequence homology and phylogenetic tree based on <em>pld</em> and <em>cp40</em> directed the observation that 2J-L is related to Mexican strain MEX29 and MEX25. Phylogenetic results agreed on the idea that strains biovar <em>ovis</em> and biovar <em>equi</em> are groupings on different clades. Finally, results indicate that Mexican strains are more similar among strains isolated from the same host type, without geography distance influence. <strong>Implications.</strong> The analysis pointed out that both genes conserve their sequences in comparison with Mexican and international strains, which encourages the research continuity for vaccine and diagnostic tools development using proteins PLD and CP40 as antigen targets. <strong>Conclusions.</strong> The complete <em>pld</em> and <em>cp40</em> genes from Mexican isolate 2J-L were amplified, cloned and analyzed; important virulence factors from <em>Corynebacterium pseudotuberculosis</em>.</p>

Bioinformatic Approach of B and T Cell Epitopes of PLD and CP40 Proteins of Corynebacterium pseudotuberculosis ovis Mexican Isolate 2J-L towards a Peptide-Based Vaccine

“…The published PLD 2J-L (accession: UYI58181) and CP40 2J-L (accession: UYI58182) protein sequences were retrieved from the National Center for Biotechnology Information (NCBI) http://www.ncbi.nlm.nih.gov/ (Accessed on 6 March 2023). Proteins belong to isolate 2J-L of Corynebacterium pseudotuberculosis biovar ovis obtained from a clinical case of Caseous Lymphadenitis in a sheep from the State of Jalisco, Mexico [23]. The Phyre2 web portal was used for modeling protein structures and producing structures in PDB format, which is necessary for conformational epitope prediction [50].…”

“…Previously, the complete sequence of the pld (GenBank accession number: OL347711) and cp40 (GenBank accession number: OL347712) genes of Corynebacterium pseudotuberculosis ovis isolate 2J-L, obtained from Jalisco State, Mexico, was reported [ 23 ]. Since those virulence factors have been studied as promising vaccine antigens, this work aimed to in silico identify B and T cell epitopes of PLD 2J-L and CP40 2J-L to determine possible peptides for a vaccine design that combines the specific components that allow effective immune system activation.…”