Treponema pallidum infection in subcutaneous polyethylene chambers in rabbits

Tight, Robert R.; Perkins, Robert L.

doi:10.1128/iai.13.6.1606-1612.1976

Cited by 8 publications

(1 citation statement)

References 11 publications

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“…Preparation of antiserum. Antiserum was raised in a 2-month-old New Zealand White rabbit by methods described previously (9,28). C. jejuni M275 glycine-hydrochloride extracts were separated by SDS-PAGE and stained with Coomassie brilliant blue R250.…”

Section: Methodsmentioning

confidence: 99%

Molecular characterization of a Campylobacter jejuni 29-kilodalton periplasmic binding protein

1996

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Campylobacter jejuni, a gram-negative, microaerophilic, spiral bacterium, is a common cause of human gastrointestinal disease. Although investigators commonly use C. jejuni glycine-hydrochloride extracts in assays to determine the products that promote the binding of the organism to eukaryotic cells, the proteins contained within these extracts remain ill defined. Characterization of these proteins will provide a better understanding of C. jejuni gene regulation and organization. An antiserum was raised against a C. jejuni 29-kDa gel-purified protein detected in glycine-hydrochloride extracts. This antiserum was used to screen an expression library of C. jejuni. A reactive clone that contained an open reading frame of 256 amino acids was identified. The cloned gene was transcribed and translated, and the product was exported to the periplasmic space in Escherichia coli XL1-Blue. The translated C. jejuni product, designated P29, exhibited significant similarity to the histidine and lysine-arginine-ornithine periplasmic binding proteins (HisJ and LAO, respectively) of Salmonella typhimurium. The C. jejuni gene encoding the P29 protein complemented an S. typhimurium HisJ mutant but not a LAO mutant when provided in trans. These data suggest that the C. jejuni gene encoding the P29 protein is a homolog of the S. typhimurium hisJ gene. Campylobacter jejuni is frequently isolated from individuals suffering from acute gastrointestinal disease (2). Infection with C. jejuni is often acquired by ingestion of undercooked chicken, unpasteurized milk, or untreated water. C. jejuni infection is characterized by the sudden onset of fever, abdominal cramps, and diarrhea with blood and leukocytes. Despite the recognition of C. jejuni as a common cause of gastrointestinal disease, the pathogenic mechanisms associated with C. jejuni gastrointestinal disease are ill defined. Potential virulence determinants include cytotoxins and the products that mediate the binding of the organism to (3, 7, 11, 13, 23) and entry into (12, 13, 17, 21) eukaryotic cells (19, 32). A number of investigators have used outer membrane protein and glycine-hydrochloride extracts in ligand-binding assays to identify the bacterial products that promote the binding of C. jejuni to eukaryotic cells (4, 6). Products proposed to facilitate binding range in molecular mass from 26 to 30 kDa (6). Although the treatment of C. jejuni with glycine-hydrochloride is thought to result in the extraction of cell surface and periplasmic proteins (18), the components of the glycine-hydrochloride extracts remain only partially characterized. To define one component of C. jejuni glycine-hydrochloride extracts, a rabbit antiserum was raised against a protein with an apparent molecular mass of 29 kDa detected in the extracts. This rabbit anti-29-kDa serum was then used to screen a C. jejuni expression library. A C. jejuni gene that encodes a protein of 256 amino acids with a calculated molecular mass of 28,531 Da (P29) was cloned. The deduced amino acid sequence of this gene exhi...

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Section: Methodsmentioning

confidence: 99%