Impact of Salmonella FimH Gene Insertion on Lactobacillus plantarum Growth and Expression for Oral Vaccine Development
DOI:
https://doi.org/10.52783/jns.v14.3237Keywords:
N\AAbstract
Salmonellosis poses a major threat to poultry health and food safety, driving the need for more effective and sustainable vaccines. In this study, we developed a recombinant Lactobacillus plantarum-based oral vaccine expressing the Salmonella FimH antigen to induce mucosal and systemic immunity in poultry. The pTRK892 shuttle vector was engineered to include a constitutive promoter (Ppgm1), a signal peptide (SP1), and anchoring motifs for efficient antigen expression and secretion. The recombinant plasmids (pTRK722 and pTRK722FimH Histag) were successfully transformed into L. plantarum, confirmed by PCR, restriction mapping, and sequencing. Western blot analysis verified the expression and secretion of the FimH fusion protein. Growth curve analysis revealed a moderate metabolic burden on recombinant strain, reflected in a slightly reduced growth rate compared to the control. This recombinant vaccine platform offers a safe, cost-effective, and scalable alternative for poultry immunization, with the potential to reduce Salmonella colonization and transmission, ultimately enhancing food safety and public health.
Downloads
Metrics
References
Angulo, F.J., Johnson, K.R., Tauxe, R.V., and Cohen, M.L., 2000. Origins and consequences of antimicrobial-resistant nontyphoidal Salmonella: Implications for the use of fluoroquinolones in food animals. Microbial Drug Resistance, 6(1), pp.77-83.
Barrow, P.A. and Lovell, M.A., 1991. Experimental infection of egg‐laying hens with Salmonella enteritidis phage type 4. Avian Pathology, 20(2), pp.335-348.
De Buck, J.D., Immerseel, F.V., Haesebrouck, F., and Ducatelle, R., 2004. Effect of type 1 fimbriae of Salmonella enterica serotype Enteritidis on bacteremia and reproductive tract infection in laying hens. Avian Pathology, 33(3), pp.314-320.
Duong, T., Miller, M.J., Barrangou, R., Azcarate‐Peril, M.A., and Klaenhammer, T.R., 2011. Construction of vectors for inducible and constitutive gene expression in Lactobacillus species. Microbial Biotechnology, 4, pp.357-367.
Hase, K., Kawano, K., Nochi, T., Pontes, G.S., Fukuda, S., Ebisawa, M., Kadokura, K., Tobe,
T., Fujimura, Y., Kawano, S., Yabashi, A., Waguri, S., Nakato, G., Kimura, S., Murakami, T.,
Iimura, M., Hamura, K., Fukuoka, S.-I., Lowe, A.W., Itoh, K., Kiyono, H., and Ohno, H., 2009. Uptake through glycoprotein 2 of FimH+ bacteria by M-cells initiates mucosal immune response. Nature, 462(7270), pp.226–230.
Kisiela, D., Kramer, J.J., Tchesnokova, V., Aprikian, P., Yarov-Yarovoy, V., Clegg, S., and Sokurenko, E.V., 2011. Allosteric catch bond properties of the Serovar Typhimurium FimH adhesin from Salmonella enterica. Journal of Biological Chemistry, 286(44), pp.3813638147.
Mathiesen, G., Sveen, A., Piard, J.C., Axelsson, L., and Eijsink, V.G.H., 2008. Heterologous protein secretion by Lactobacillus plantarum using homologous signal peptides. Journal of Applied Microbiology, 105(1), pp.215-226.
McSorley, S.J. and Jenkins, M.K., 2000. Antibody is required for protection against virulent but not attenuated Salmonella enterica serovar Typhimurium. Infection and Immunity, 68(6), pp.3344-3348.
Mukkur, T.K.S., McDowell, G.H., Stocker, B.A.D., and Lascelles, A.K., 1987. Protection against experimental salmonellosis in mice and sheep by immunization with aromaticdependent Salmonella typhimurium. Journal of Medical Microbiology, 24(1), pp.11-19.
Nouaille, S., Ribeiro, L.A., Miyoshi, A., Pontes, D., Le Loir, Y., Oliveira, S.C., Langella, P., and Azevedo, V., 2003. Heterologous protein production and delivery systems in Lactococcus lactis. Genetic Engineering Research, 2, pp.102-111.
Saini, S., Pearl, J.A., and Rao, C.V., 2009. Role of FimW, FimY, and FimZ in regulating the expression of type I fimbriae in Salmonella enterica serovar Typhimurium. Journal of Bacteriology, 191, pp.3003-3010.
Sorvig, E., Gronqvist, S., Naterstad, K., Mathiesen, G., Eijsink, V.G., and Axelsson, L., 2003.
Construction of vectors for inducible gene expression in Lactobacillus sakei and L. plantarum. FEMS Microbiology Letters, 22, pp.119-126.
Steidler, L., 2002. In situ delivery of cytokines by genetically engineered Lactococcus lactis.
Antonie Van Leeuwenhoek, 82, pp.323-331.
Sztein, M.B., Wasserman, S.S., Tacket, C.O., Edelman, R., Hone, D., Lindberg, A.A., and Levine, M.M., 1994. Cytokine production patterns and lymphoproliferative responses in volunteers orally immunized with attenuated vaccine strains of Salmonella typhi. Journal of Infectious Diseases, 170(6), pp.1508-1517.
Tizard, I.R., 2004. Veterinary Immunology (7th ed.). Saunders Publication, Philadelphia, United States.
van Rooijen, R.J., and de Vos, W.M., 1990. Molecular cloning, transcriptional analysis, and nucleotide sequence of lacR, a gene encoding the repressor of the lactose phosphotransferase system in Lactococcus lactis. Journal of Biological Chemistry, 265, pp.18499-18503.
Voskuil, M.I., and Chambliss, G.H., 1998. The -16 region of Bacillus subtilis and other grampositive bacterial promoters. Nucleic Acids Research, 26, pp.3584-3590.
Wells, J.M., Wilson, P.W., Norton, P.M., Gasson, M.J., and Le Page, R.W., 1993. Lactococcus lactis: High-level expression of tetanus toxin fragment C and protection against lethal challenge. Molecular Microbiology, 8, pp.1155-1162.
Yeh, K.S., Tinker, J.K., and Clegg, S., 2002. FimZ binds the Salmonella typhimurium fimA promoter region and may regulate its own expression with FimY. Microbiology and Immunology, 46, pp.1-10.
Downloads
Published
How to Cite
Issue
Section
License
This work is licensed under a Creative Commons Attribution 4.0 International License.
You are free to:
- Share — copy and redistribute the material in any medium or format
- Adapt — remix, transform, and build upon the material for any purpose, even commercially.
Terms:
- Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.
- No additional restrictions — You may not apply legal terms or technological measures that legally restrict others from doing anything the license permits.
