Influence of heat-labile serum components in the presence of OmpA on the outer membrane of Salmonella gallinarum
| Autor: | Daniel Martínez-Gómez, Yolanda López-Vidal, Antonio Verdugo-Rodríguez, L Huerta-Ascencio, X D Vega-Manriquez |
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| Rok vydání: | 2015 |
| Předmět: |
Serum
0301 basic medicine Salmonella Hot Temperature Salmonella Gallinarum medicine.disease_cause Polymerase Chain Reaction Biochemistry Microbiology law.invention 03 medical and health sciences Antigen law Genetics medicine Animals Molecular Biology Gene Incubation Polymerase chain reaction biology General Medicine bacterial infections and mycoses biology.organism_classification 030104 developmental biology bacteria Electrophoresis Polyacrylamide Gel Bacterial outer membrane Chickens Bacteria Bacterial Outer Membrane Proteins |
| Zdroj: | Archives of Microbiology. 198:161-169 |
| ISSN: | 1432-072X 0302-8933 |
| Popis: | Salmonella gallinarum is the causative agent of fowl typhoid. Being a Gram-negative bacteria, its outer membrane proteins (OMP) can be regulated by different microenvironments. S. gallinarum was cultured under the following conditions: nutrient broth (NB), NB supplemented with serum from specific pathogen-free birds (NBS) and NB with serum incubated at 56 °C prior to incubation with the bacteria (NBSD); OMP were subsequently extracted. Several changes were observed in the apparent expression of OMP, mainly a decrease in an OMP with a size of 30 kDa, approximately, under the NBS condition. In contrast, the same event was not observed in NB and NBSD when using one- and two-dimensional polyacrylamide gels (SDS-PAGE). Using the OMP with a size of 30 kDa, approximately, as antigen in indirect ELISA, we were able to differentiate serum from healthy and vaccinated birds, as well as birds infected with S. gallinarum and S. enteritidis. The amino-terminal of this protein was sequenced, showing 100 % identity with OmpA of S. typhimurium. Subsequently, we designed primers to amplify the gene by PCR. The partial sequence of the amplified gene showed 100 % identity with OmpA of S. gallinarum. (1) Heat-labile serum components influence the presence of OmpA in the OM of S. gallinarum; (2) by the way of ELISA, OmpA allows to specifically differentiate healthy from diseased birds. |
| Databáze: | OpenAIRE |
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