Submitted to: Clinical and Vaccine Immunology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/19/2006
Publication Date: 1/3/2007
Citation: Li, L., Munir, S., Bannantine, J.P., Sreevatsan, S., Kanjilal, S., Kapur, V. 2007. Rapid Expression of Mycobacterium avium subsp. paratuberculosis Recombinant Proteins for Antigen Discovery. Clinical and Vaccine Immunology. 14(1):102-105.
Interpretive Summary: Making recombinant proteins is labor intensive. Therefore, obtaining thousands of recombinant proteins for antigen studies is virtually impossible. Nonetheless, this systematic approach is necessary to unequivocally identify the strongest antigens present in Mycobacterium avium subspecies paratuberculosis. This communication describes our efforts to produce recombinant proteins in a high-throughput manner. The novel in vitro transcription and translation system combined with the transcriptionally active PCR is one way to obtain hundreds of recombinant proteins quickly. We introduce this technique and demonstrate its utility with two selected Mycobacterium avium subspecies paratuberculosis proteins.
Technical Abstract: Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne’s disease, a chronic granulomatous enteritis of ruminants and other species. Detection of infection in animals is hampered by the lack of sensitive and specific diagnostic assays. We here describe an approach that utilizes translationally active PCR fragments for the rapid in vitro transcription and translation of recombinant proteins for antigen discovery in M. paratuberculosis. Using this approach, our investigations show that the MAP1272c protein selectively reacts with sera from Johne’s disease positive cattle and represents an antigen of potential utility in M. paratuberculosis immunodiagnostics.