A novel antigen from Anaplasma marginale: characterization, expression and preliminary evaluation of the recombinant protein |
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Authors: | Hope Michelle Riding George Menzies Moira Willadsen Peter |
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Affiliation: | Cooperative Research Center for Vaccine Technology and CSIRO Livestock Industries, Queensland Biosciences Precinct, 306 Carmody Road, QLD 4067, St. Lucia, Australia. shelly.hope@csiro.au |
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Abstract: | Through a process of protein fractionation and vaccination we previously identified four native antigens that confer a degree of protection against challenge with Anaplasma marginale. One of these, Ana 29 has been successfully cloned and sequenced using degenerate primers designed to N-terminal and internal peptide sequences. The full-length gene codes for a protein with a theoretical molecular weight of 27 kDa and pI 8.6. The sequence is highly conserved, showing 99% identity between two Australian and an American isolate of A. marginale. The gene sequences from these isolates also share 99% identity with the strain of Anaplasma centrale used in the commercial Australian vaccine. Protein prediction algorithms suggest the native protein is an integral membrane protein. This protein has been over-expressed and purified from Escherichia coli and used in vaccination trials in cattle using two adjuvants. The initial results from the trial show a significant level of protection was obtained with one adjuvant; in comparison, the second adjuvant slightly aggravated the disease. Preliminary data suggests a good correlation between the induction of an IgG2 response and protection. |
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