Anti-cytoskeleton Immunoscreening of Trypanosoma brucei Expression Library Reveals Novel Immunogenic Conserved Putative Proteins

The overall shape of the trypanosome is defined by an internal cytoskeleton consisting of a network of microtubules that are cross-linked both to each other and the inner face of the plasma membrane. However, the total complement and identity of the trypanosome cytoskeleton proteins are not yet fully determined despite the fact that some of them may be good targets for diagnostics, drugs, and/or vaccine discovery. Therefore, in this study, rabbit anti-Trypanosoma brucei detergent-insoluble cytoskeleton sera were produced in vivo and used to probe a T. brucei expression library. The picked plaques were made clonal by a series of library screenings followed by PCR amplification, cDNA sequencing, and identification of the proteins coded by these sequences using BLAST. The previously well-known cytoskeleton proteins (paraflagella rod protein and histone H2B), putative cytoskeleton proteins (Dynein light chain and nucleoporin), conserved hypothetical protein (Tb10.61.2430), and novel cytoskeleton protein coding cDNA sequences (not in the sequenced and published T. brucei genome) were identified in this study. This approach is therefore usable in the search for novel proteins whose utility in the design and development of diagnostics, drugs, and/or vaccines can further be studied.