Cloning of Brucella abortus gene and characterization of expressed 26-kilodalton periplasmic protein: potential use for diagnosis.
Brucella spp. are the causative agents of brucellosis in many different hosts, including humans. Most of the serological methods of diagnosis are based on the detection of antilipopolysaccharide antibodies, which makes the differentiation of vaccinated animals from infected animals difficult. By using molecular biology techniques, a gene that encodes a 26-kDa protein (BP26) was isolated from a Brucella abortus S19 genome lambda gt11 library. This protein is in the periplasm of B. abortus and in transformed Escherichia coli. It is exported to the periplasm via a preprotein of 29 kDa with a signal sequence of 28 amino acids. The nucleotide and amino acid sequences of this gene and protein did not show any similarity with those of previously sequenced genes. The use of this protein in Western blotting allowed the differentiation between vaccinated bovines from infected bovines and the detection of infected rams: on the other hand, sera from human patients with active brucellosis were positive, while sera from human patients with chronic brucellosis or without clinical signs were nonreactive. BP26 might be of value as an antigen for serological diagnosis of brucellosis in different mammals.
Rossetti
O L
OL
Instituto de Biotecnología, Centro de Investigación en Ciencias Veterinarias, Buenos Aires, Argentina. ROL@bminta.edu.ar
Arese
A I
AI
Boschiroli
M L
ML
Cravero
S L
SL
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D002003:Brucella abortus / Q000235:genetics*; D002006:Brucellosis / Q000175:diagnosis* / Q000276:immunology / Q000662:veterinary; D002007:Brucellosis, Bovine / Q000175:diagnosis* / Q000276:immunology; D002417:Cattle; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D005260:Female; D005798:Genes, Bacterial; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011247:Pregnancy; D012698:Serologic Tests; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis / Q000276:immunology
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial