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AssigneeKansas University Research Foundation
InventorsT.G. Nagaraja, George C. Stewart, Sanjeev K. Narayanan, M. M. Chengappa
Estimating the term…
The F. necrophorum gene expressing leukotoxin was sequenced and cloned. The leukotoxin open reading frame (lktA) is part of a multi-gene operon containing 9,726 bp, and encoding a protein containing 3,241 amino acids with an overall molecular weight of 335,956 daltons. The protein encoded by the gene was truncated into five polypeptides having overlapping regions by truncating the full length gene into five different sections and amplifying, expressing, and recovering the protein encoded by each of these sections. Additionally, a region upstream of the gene was sequenced and the polypeptide encoded by that nucleotide sequence was purified and isolated. These polypeptides along with the full length protein are then tested to determine their immunogenicity and protective immunity in comparison to the efficacy of immunization conferred by inactivated native leukotoxin in F. necrophorum culture supernatant.
An isolated polypeptide having an amino acid sequence having at least about 50% sequence homology with a sequence selected from the group consisting of SEQ ID Nos. 2-7.
An isolated nucleotide sequence having a nucleotide sequence having at least about 50% sequence homology with a sequence selected from the group consisting of SEQ ID Nos. 8-14.
An expression vector containing a nucleotide sequence having at least about 50% sequence homology with a sequence selected from the group consisting of SEQ ID Nos. 8-14.
A vaccine comprising a polypeptide having a sequence having at least about 50% sequence homology with a sequence selected from the group consisting of SEQ ID Nos 2-7.
A recombinantly derived polypeptide having a sequence having at least about 50% sequence homology with a sequence selected from the group consisting of SEQ ID Nos 1-7.
A method of preparing a vaccine which confers effective immunity against infection caused by F. necrophorum comprising the steps of:
A method of immunizing an animal against liver abscesses caused by F. necrophorum comprising the steps of:
A recombinant polypeptide sequence which is recognized by anti-native leukotoxin antibodies in a western blot analysis.
A recombinant polypeptide sequence whose antisera neutralizes activity of native leukotoxin against bovine polymorphonuclear leukocytes, said sequence having at least about 50% sequence homology with a sequence selected from the group consisting of SEQ ID Nos. 1, 2 and 4.
A recombinantly derived polypeptide sequence effective in conferring protective immunity against F. necrophorum infection in mice, said sequence having at least about 50% sequence homology with a sequence selected from SEQ ID Nos. 9 and 12.
Dependencies are inferred from the claim text.