Résumé
Lactobacillus casei 393 was selected as a bacterial carrier for the expression of Porcine Parvovirus (PPV) protective antigen VP2 protein. The gene encoding PPV VP2 protein was cloned into the Lactobacillus casei surface expression vector pPG, and then the constructed recombinant vector pPG-VP2 was electrotransformed into Lactobacillus casei 393 generating the recombinant system pPG-VP2/L. casei393 expressing PPV VP2 protein. The recombinant strain was induced by 2% Lactose in MRS and about 74kD protein was detected with SDS-PAGE. The result of Western-blot indicated that the expressed protein possessed the antigenic specificity which could be recognized by mouse anti-PPV serum. The indirect immunofluorescent test showed that the expressed protein was secreted on the cell surface Lactobacillus casei.
Sujets)
Animaux , Antigènes viraux , Génétique , Métabolisme , Technique de Western , Protéines de capside , Génétique , Métabolisme , Membrane cellulaire , Métabolisme , Électrophorèse sur gel de polyacrylamide , Technique d'immunofluorescence , Lacticaseibacillus casei , Génétique , Métabolisme , Parvovirus porcin , Génétique , Métabolisme , Plasmides , Génétique , Protéines recombinantes , Métabolisme , Suidae , Virologie , Transformation génétique , Protéines virales , Génétique , MétabolismeRésumé
<p><b>UNLABELLED</b>The gene encoding classical swine fever virus (CSFV) T cell epitope E290 peptide was synthesized by PCR, cloned into the expression vector pPG-VP2 and named pPG-VP2-E290. The recombinant plasmid was electrotransformed into Lactobacillus casei 393 generating pPG-VP2-E290/L. casei 393. Specific anti-CSFV E290 peptide immunoglobulin G (IgG) antibody was detected by indirect ELISA in the serum of BALB/c mice and rabbits immunized with recombinant strain by oral administration. The CTL of E290 was analyzed with lymphocytes taken from the immunized mice, and the immunized rabbits were attacked with CSFV to validate the protective function of E290 antibody induced.</p><p><b>RESULT</b>The recombinant expression system constructed with L. casei 393 in this study show a good immunization property and could elicit the mice and rabbits to produce high anti-E290 antibody levels. Furthermore, E290 peptide antibody could elicit specific CTL response, and restrain attack of CSFV to rabbits.</p>