Figure 2: Expression of P97R1N in S. enterica serovar Choleraesuis C501. Vaccine strain C501 (P97R1N) and vector control strain C501 (pYA3493) were cultured in LB broth at 37°C. Total cells and concentrated culture supernatants were subjected to SDS-PAGE analysis, and P97R1N was detected by immunoblotting with anti-M. hyopneumoniae antibody. (a) Coomassie brilliant blue-stained gel analysis. M, protein ladder; lane 1, total cell extracts of C501 (pYA97R1N); lane 2, concentrated supernatants of C501 (pYA97R1N); lane 3, C501 (pYA3493); (b) Immunoblot analysis. M, protein ladder; lane 1, total cell extracts of C501 (P97R1N); lane 2, concentrated supernatants of C501 (pYA97R1N); lane 3, C501 (pYA3493). (c) Analysis of P97R1N stable epression from the recombinant strain C501 (pYAP97R1N) by SDS-PAGE. Lane 1-9, second passage to ten passage from C501 (pYAP97R1N), lane 10, C501 (pYA3493).