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| Peptide Synthesis | Protein samples were separated by SDS-PAGE on 12.5% polyacrylamide gels and stained with Coomassie Blue. For western blot analysis, the immunogenic peptide SCQKDRNDFAFPQDV was synthesized by GenScript. Mice were immunized with this peptide in the same laboratory (GenScript) to produce polyclonal anti-FeIFN-α7 primary antibody. Gels were transferred onto a nitrocellulose membrane and FeIFN-α7 and FeIFN-α7xArg were detected using the mouse polyclonal anti-FeIFN-α7 as the primary antibody and HRP-conjugated anti-mouse IgG (DAKO, Copenhagen, Denmark) as the secondary antibody. For 3,3′-diaminobenzidine (DAB) staining, the gel was immersed in a 9 mg ml−1 DAB aqueous solution (Sigma) with 10 μl of 2.6 M hydrogen peroxide for 2 min. Page ruler prestained protein ladder (Thermo Scientific, Rockford, IL USA) was used as the molecular weight marker. | Get A Quote |
Feline interferons (FeIFNs) are cytokines with antiviral, antitumor and immunomodulatory functions used as therapeutic agents in a variety of veterinary diseases. In this work, FeIFN-α7 and FeIFN-α7xArg containing eight residues of arginine were expressed in Sf9 cells and insect larvae. At 4 days post-infection (dpi), the concentrations of FeIFN-α7 and FeIFN-α7xArg in suspension culture were (1.28 ± 0.15) × 106 U ml−1 and (1.3 ± 0.2) × 106 U ml−1 respectively. The maximum expression levels of FeIFN-α7 and FeIFN-α7xArg were (3.7 ± 0.2) × 106 U ml−1 and (3.5 ± 0.4) × 106 U ml−1 at 2 dpi in Rachiplusia nu larvae and (1.1 ± 0.2) × 106 U ml−1and (1.0 ±... More
