Start Date: Dec 01, 2006
End Date: Nov 30, 2011
Plasmid constructions will be based on vector backbones that have shown the ability to express mycobacterial antigens invitro, producing adequate levels of recombinant proteins for further analysis. Plasmids will be constructed using gene synthesis by commercial companies. After construction of synthetic genes in expression vectors, these proteins will be produced in E. coli. The recombinant proteins will be tested in experimentally infected cattle for evaluation of T-cell and B-cell responses to the purified proteins. The Bovigam IFN-gamma procedure will be standardized based on the results of the in vitro tests. Different peptide cocktail formulations (including stabilizers for a future in-tube stimulation) will be evaluated.