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Purification and partial characterization of type 3 fimbriae from Salmonella typhimurium var. copenhagen
H Stolpe1, S Grund, W Schröder
1Institut für Mikrobiologie, Elektronenmikroskopie, Fachbereich Veterinärmedizin, Freie Universität Berlin, Germany.
Abstract:
Aggregative thin fimbriae from a pigeon pathogen, Salmonella typhimurium var. copenhagen (STMVC) Mö 8 were isolated and purified. These fimbriae remained associated with the cells even after attempts to separate them from blended cells by centrifugation. After purification, fimbriae and little cell fractions were polymerized in formic acid and then analyzed by SDS-PAGE. This pretreatment resulted in the appearance of a main protein band of 17 kDa. The N-terminal amino acid sequence of 19 residues of purified 17 kDa protein showed considerable homology with the N-terminal sequence of thin fimbriae of S. enteritidis. Native fimbriae on whole cells were specifically labelled with immune serum raised to the purified fimbriae. This immune serum also reacted with the denatured 17 kDa protein in Western blots. The polyclonal immune serum did not cross-react with the type-1 fimbriae produced by STMVC.
Insights
Researchers isolated and purified aggregative thin fimbriae from Salmonella typhimurium var. copenhagen (STMVC), a pigeon pathogen. The study identified a 17 kDa protein component homologous to S. enteritidis fimbriae, aiding Salmonella research.
Area of Science:
- Microbiology
- Bacteriology
- Protein Chemistry
Background:
- Salmonella typhimurium var. copenhagen (STMVC) is a significant avian pathogen.
- Fimbriae, hair-like appendages on bacterial surfaces, play crucial roles in adhesion and pathogenesis.
- Understanding the specific fimbrial structures of STMVC is essential for developing targeted control strategies.
Purpose of the Study:
- To isolate and purify aggregative thin fimbriae from Salmonella typhimurium var. copenhagen (STMVC).
- To characterize the protein components of these fimbriae, including their molecular weight and N-terminal sequence.
- To generate specific antibodies for identifying and differentiating these fimbriae from other Salmonella fimbriae.
Main Methods:
- Isolation and purification of fimbriae from bacterial cells.
- Polymerization of fimbriae and cell fractions in formic acid.
- Analysis using SDS-PAGE to determine protein band sizes.
- N-terminal amino acid sequencing of purified proteins.
- Immune serum production and characterization using Western blotting and whole-cell labeling.
Main Results:
- Aggregative thin fimbriae were successfully isolated and purified from STMVC.
- A predominant 17 kDa protein band was identified after formic acid polymerization and SDS-PAGE.
- The N-terminal sequence of the 17 kDa protein exhibited homology with thin fimbriae from Salmonella enteritidis.
- Specific polyclonal immune serum recognized native and denatured fimbriae but did not cross-react with type-1 fimbriae of STMVC.
Conclusions:
- The study successfully characterized the aggregative thin fimbriae of Salmonella typhimurium var. copenhagen.
- A 17 kDa protein is a major component of these fimbriae, sharing homology with fimbriae from other Salmonella species.
- The developed immune serum provides a specific tool for identifying these fimbriae, distinguishing them from type-1 fimbriae.