Volume 32, Issue 8 (November 2021)
Studies in Medical Sciences 2021, 32(8): 572-580 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Saki Hosseini M, Doosti A, Pezeshki M. GENERATION OF A GENE CONSTRUCT TO SIPA GENE DELETION OF SALMONELLA TYPHIMURIUM. Studies in Medical Sciences 2021; 32 (8) :572-580
URL: http://umj.umsu.ac.ir/article-1-5412-en.html
URL: http://umj.umsu.ac.ir/article-1-5412-en.html
Assistant Professor, Department of Genetics, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran (Corresponding Author) , m.saki.hoseini@gmail.com
Abstract: (2727 Views)
Background & Aims: Salmonella Typhimurium is a negative-gram, non-spore, free capsule, moving bacteria with Trish Perry flagella. Salmonella is the most common cause of food poisoning. The ability to enter and survive in host cells is the condition for pathogenic Salmonella species. Proteins of invasive Salmonella are transferred to the host cells by bacteria. This study was performed for generation of a gene construct to SipA gene deletion of Salmonella Typhimurium and its cloning in E.Coli bacteria.
Materials & Methods: This laboratory study was conducted in biotechnology research center of Islamic Azad University Shahrekord Branch from September 2017 to May 2018. In this study, 5' and 3' sequence of SipA gene was amplified by the specific primers and PCR method. Then, each of these sequences was cloned by the T/A cloning method in pGEM-Teasy vector and then was transformed into E.Coli bacteria. Using the PCR method, the part related to each region was amplified and confirmed. The final confirmation of the produced construct was performed by the Xbal and Xhol enzymes.
Results: The results indicated the successful cloning of the target gene in E.Coli and generation of a gene construct with a length of the 1520 bp. Also, pET32 vector with a length of 5900 bp was the best vector for the admission construct.
Conclusion: Based on the results, it seems that by inserting a gene, the damaged gene can be deleted and it can be used in the future research as a gene vaccine against Salmonellosis. Also, the target gene can be deleted with electroporation method and transferred into Salmonella Typhimurium, and due to the similarities between upstream and downstream gene sequences of sipA and Kan genes, homologous recombination between these two genes can occur and pathogenic genes will be removed.
Materials & Methods: This laboratory study was conducted in biotechnology research center of Islamic Azad University Shahrekord Branch from September 2017 to May 2018. In this study, 5' and 3' sequence of SipA gene was amplified by the specific primers and PCR method. Then, each of these sequences was cloned by the T/A cloning method in pGEM-Teasy vector and then was transformed into E.Coli bacteria. Using the PCR method, the part related to each region was amplified and confirmed. The final confirmation of the produced construct was performed by the Xbal and Xhol enzymes.
Results: The results indicated the successful cloning of the target gene in E.Coli and generation of a gene construct with a length of the 1520 bp. Also, pET32 vector with a length of 5900 bp was the best vector for the admission construct.
Conclusion: Based on the results, it seems that by inserting a gene, the damaged gene can be deleted and it can be used in the future research as a gene vaccine against Salmonellosis. Also, the target gene can be deleted with electroporation method and transferred into Salmonella Typhimurium, and due to the similarities between upstream and downstream gene sequences of sipA and Kan genes, homologous recombination between these two genes can occur and pathogenic genes will be removed.
References
1. Ranjbar R, Sarshar M. Genetic diversity of clinical strains of Salmonella enteric serovar Typhimurium. J Mil Med 2012;14(2):143-7. [Google Scholar]
2. Xiaojuan Y, Jiahui H, Qingping Wu, Jumei Zh, Shengrong Liu, Weipeng G, et al. Prevalence, antimicrobial resistance and genetic diversity of Samonella isolated from retail ready- to- eat foods in china. Food Control 2016;60:50-6. [DOI:10.1016/j.foodcont.2015.07.019]
3. Ranjbar R, Giammanco GM, Farshad S, Owlia P, Aleo A, Mammina C. Serotypes, antibiotic resistance and class 1integrons in Salmonella isolates from pediatric cases of enteritis in Tehran, Iran. Foodborne Pathog Dis 2011;8(4):547-53. [DOI:10.1089/fpd.2010.0736] [PMID]
4. Tennant SM, Levine MM. Live attenuated vaccines for invasive Salmonella infections. Journal: Vaccine. Vaccine 2015;33:C36-41. [DOI:10.1016/j.vaccine.2015.04.029] [PMID] [PMCID]
5. Ranjbar R, Giammanco GM, Aleo A, Plano MR, Naghoni A, Owlia P, et al. Characterization of the first extendedspectrum b-lactamase producing nontyphoidal Salmonella strains isolated in Tehran, Iran. Foodborne Pathog Dis 2010;7(1):91-5. [DOI:10.1089/fpd.2009.0382] [PMID]
6. Naghoni A, Ranjbar R, Tabaraie B, Farshad S, Owlia P, Safiri Z, et al. High prevalence of integron-mediated resistance in clinical isolates of Salmonella enteric. Jpn J Infect Dis 2010;63(6):417-21. [DOI:10.7883/yoken.63.417] [PMID]
7. Skjolaas KA, Burkey TE, Dritz SS, Minton JE. Effects of Salmonella enterica Serovars Typhimurium (ST) and Choleraesuis (SC) on chemokine and cytokine expression in swine ileum and jejunal epithelial cells. Vet Immunol Immunopathol 2006;111(3-4):199 209. [DOI:10.1016/j.vetimm.2006.01.002] [PMID]
8. Ranjbar R, Soltan dalal MM, Talebi M, Pourshafie MR. Increased isolation and characterization of Shigella sonnei obtained from hospitalized children in Tehran, Iran. J Health Popul Nutr 2008;26(4):426-30. [DOI:10.3329/jhpn.v26i4.1884] [PMID] [PMCID]
9. Jianghui Zhu, Yao Bai, Yeru Wang, Xiaoyu Song, Shenghui Cui, Haibin Xu, et al. A risk assessment of salmonellosis linked to chicken meals prepared in households of china. Food control;79:279-87 [DOI:10.1016/j.foodcont.2017.04.003]
10. Farré MR, Sánchez DO, Varela CA, Sanahuja MS, Recasens AR, Jové JP. Aspectos epidemiologicos and carga asistencial de gastroenteritis agudas por Campylobacter and salmonella. Medicina Clínica 2015;145(7):294-7. [DOI:10.1016/j.medcli.2014.11.016] [PMID]
11. Desin TS, Koster W, Potter AA. Salmonella vaccines in poultry: past, present and future. J Expert Rev Vaccines 2013; 12(1): 87-96. [DOI:10.1586/erv.12.138] [PMID]
12. Abernaty J, Corkill C, Hinojosa C, Li X, Zhou H. Deletions in the pyruvate pathway of. Salmonella Typhimurium SPI1 mediated gene expression and infectivity. J Anim Sci Biotechnol 2013; 4(1):5. 1-12. [DOI:10.1186/2049-1891-4-5] [PMID] [PMCID]
13. Abernathy J, Corkill C, Hinojosa C, Li X, Zhou H. Deletions in the pyruvate pathway of Salmonella Typhimurium alter SPI1-mediated gene expression and infectivity. J Anim Sci Biotechnol 2013;4(1):5. [DOI:10.1186/2049-1891-4-5] [PMID] [PMCID]
14. .Silva M, Song C, Nadeau WJ, Matthews JB, McCormick BA. Salmonella Typhimurium sipA induced neutrophil transepithelial migration: involvement of a PKC-α-dependent signal transduction pathway. Am J Gastrointest Liver physiol 2004; 286(6): G1024-31. [DOI:10.1152/ajpgi.00299.2003] [PMID]
15. Cooper GL, Venables LM, Woodward MJ, Hormaeche CE. Vaccination of chickenswith strain CVL30, a genetically definedSalmonella enteritidisaroA live oral vaccinecandidate. Infect Immun 1994; 62(11): 4747-54. [DOI:10.1128/iai.62.11.4747-4754.1994] [PMID] [PMCID]
16. Cerquetti MC, Gherardi MM. Vaccinationof chickens with a temperature-sensitiv mutant of Salmonella enteritidis. Vaccine 2000;18(11-12): 1140-5. [DOI:10.1016/S0264-410X(99)00377-1]
17. Taseen S Desin, Wolfgang Köster, Andrew A Potter. Salmonella vaccines in poultry: past, present and future. Expert Rev Vaccines 2013: 12(1): 87-96. [DOI:10.1586/erv.12.138] [PMID]
18. Riyaz-Ul-Hassan S, Verma V, Qazi GN. Rapid detection of salmonella by polymerase chain reaction.J Mol Cell Probes 2004; 18(5): 333-9. [DOI:10.1016/j.mcp.2004.05.003] [PMID]
19. Rosu V, Chadfield MS, Santona A, Christensen JP, Thomsen LE, Rubino S, et al. Effects of crp deletion in Salmonella enterica serotype Gallinarum. Acta Vet Scand 2007;49(1):14. [DOI:10.1186/1751-0147-49-14] [PMID] [PMCID]
20. Pochop J, Kačániová M, Hleba L, Lopašovský Ľ, Rovná K, Arpášová H. Application of Real-time PCR for Rapid Petection of Salmonella spp, Salmonellaenterica ser Typhimurium and Enteritidis in Food Samples of Animal Origin without Pre-enrichment and with Pre-enrichment. Scientific Papers Animal Science and Biotechnologies2012; 45(1): 341-5. [Google Scholar]
21. Abernaty J, Corkill C, Hinojosa C, Li X, Zhou H. Deletions in the pyruvate pathway of. Salmonella TyphimuriumSPI1-mediated gene expression and infectivity. J Anim Sci Biotechnol 2013; 4(1):5: 1-12. [DOI:10.1186/2049-1891-4-5] [PMID] [PMCID]
22. Gantois I, Ducatelle R, Timbermont L, Boyen F, Bohez L, Haesebrouck F, et al. Oral immunisation of laying henswith the live vaccine strains of TADSalmonella vac E and TAD Salmonella vac T reduces internal egg contamination withSalmonella enteritidis.Vaccine 2006; 24(37-39): 6250-5. [DOI:10.1016/j.vaccine.2006.05.070] [PMID]
23. Springer S, Lindner T, Ahrens M, Woitow G, Prandini F, Selbitz HJ. Duration of immunity induced in chickens by anattenuated live Salmonella enteritidisvaccine and an inactivated Salmonellaenteritidis/typhimuriumvaccine. Berl Munch Tierarztl Wochenschr 2011;124(3-4): 89-93. [Google Scholar]
Send email to the article author
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
