Molecular cloning of a gene encoding the immunogenic 21 kDa protein of Cowdria ruminantium

摘要: Major immunogenic polypeptides (21, 32, 40, 46, 58, 85 and 160 kDa) of Cowdria ruminantium were identified by immunoprecipitation immunoblotting. A pUC13 library C. genomic DNA was screened with hyperimmune sheep serum to identify Escherichia coli colonies which expressed genes encoding these proteins. recombinant E. colony, F5.2, containing plasmid insert 2773 bp. The cloned contained two long open reading frames (ORFs) 627 bp (complete) 831 (incomplete), both potentially proteins an N-terminal signal peptide. Deletion experiments suggested that the recognized a protein encoded ORF. ORF amplified polymerase chain reaction (PCR), subcloned high level in coli. antiserum made fusion 21 kDa all strains tested, confirming encodes native ruminantium. Similarly, sera tested from heartwater-infected animals. antigenic conservation its nature are reasons for further testing this subunit diagnostic tests.