Products/Services Used | Details | Operation |
---|---|---|
Gene Synthesis | Cloning of cat flea genes encoding candidate protective antigens and production of recombinant proteins Coding sequences for selected candidate protective anti- gens were amplified by RT-PCR using sequence-specific primers (F1-F4) or synthesized (GenScript, Hong Kong) with optimized codon usage for Escherichia coli (F5, F6) (see Datasets 2 and 3 in Additional file 2)....5 h to produce re- combinant proteins, which were purified to > 95% of total cell proteins by Ni affinity chromatography (Genscript Corporation, Piscataway, NJ, USA) as previously described [27, 54, 55] using 1 ml HisTrap FF columns mounted on an AKTA-FPLC system (GE Healthcare, Piscataway, NJ, USA) in the presence of 7 M urea lysis buffer. | Get A Quote |
Despite the abundance of the domestic cat flea, Ctenocephalides felis (Bouché, 1835) and disease risks associated with them, flea control is difficult and requires the development of new control interventions such as vaccines. In this study, a reverse vaccinology approach was designed to achieve a rational selection of cat flea candidate protective antigens.,Based on transcriptomics and proteomics data from unfed adult fleas it was possible to select more specific candidate protective antigens based on highly represented and functionally relevant proteins present in the predicted exoproteome. The protective capacity of the recombinant antigens was evaluated for the control of C. felis infestations in vaccinate... More