Cloning, Prokaryotic Soluble Expression, and Analysis of Antiviral Activity of Two Novel Feline IFN-? Proteins

Wang, Xiaona; Li, Fengsai; Han, Meijing; Jia, Shuo; Wang, Li; Qiao, Xinyuan; Jiang, Yanping; Cui, Wen; Tang, Lijie; Li, Yijing; Xu, Yi-Gang

The characteristics of the novel feline IFN-ω proteins (feIFN-ωa and feIFN-ωb) were analyzed using several online bioinformatics software programs, including the identification of potential signal peptide cleavage sites, N-glycosylation sites, O-glycosylation sites, phosphorylation sites, subcellular localization, and transmembrane regions. The results from these detailed analyses are displayed in Table 1 . In addition, we also used online software algorithms to predict antigen epitopes, hydrophobicity, and the secondary and three-dimensional structures of the feIFN-ωa and feIFN-ωb proteins. As shown in Figure Figure 3b ). The maximum hydrophobicity of feIFN-ωa was 1.92, and the minimum hydrophobicity was −2.27. The maximum hydrophobicity of feIFN-ωb was 2.27, and the minimum hydrophobicity was −2.36. The secondary structure of the two feIFN-ω proteins was predicted using SOPMA software and revealed that feIFN-ωa contained 62.24% alpha helix, 2.55% beta sheet, and 34.18% irregular curl structures (Figure 3c ), whereas feIFN-ωb contained 65.52% alpha helix, 1.97% beta sheet, and 31.53% irregular curl structures ( Figure 3d ). The three-dimensional structures of feIFN-ωa and feIFN-ωb were predicted with SWISS-MODEL software and are shown in Figure 3e ,f, respectively. (6) 84.3% extracellular 9.2% intracellular 7% mitochondrion Intracellular a signal peptide cleavage sites were analyzed by SignalP 3.0 Server at http://www.cbs.dtu.dk/services/ SignalP-3.0/; b phosphorylation sites were analyzed by NetPhos3.1 Server at http://www.cbs.dtu.dk/services/ NetPhos/; c N-glycosylation sites were analyzed by NetNGlyc1.0 at http://www.cbs.dtu.dk/services/NetNGlyc/ and O-glycosylation sites were analyzed by YinO Yang1.2 at http://www.cbs.dtu.dk/services/YinOYang/; d subcellular localization was analyzed by TargetP 1.1 Server at http://www.cbs.dtu.dk/services/TargetP; e transmembrane region was analyzed by TMHMM Server v. 2.0 at http://www.cbs.dtu.dk/services/TMHMM/.