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

We first collected splenic lymphocytes from cats infected with VSV and treated with poly(I:C) simultaneously. We then performed RT-PCR assays using a pair of degenerate primers to identify two genes encoding feIFN-ω, referred to as feIFN-ωa and feIFN-ωb ( Figure 1a) . These two novel genes were deposited in the GenBank with accession numbers MK682680 and MK682681. Following nucleotide sequence homology analysis, we found that the feIFN-ωa gene (MK682680), with a size of 591 bp, shared a maximum nucleotide sequence homology of 91.88% (91.6% amino acid homology) with the 13 known subtypes of feIFN-ω (feIFN-ω1 to feIFN-ω13) genes published in the GenBank (Figure 1c ). The feIFN-ωb gene (MK682681), with a size of 612 bp, shared a maximum sequence homology of 90.20% (89.4% amino acid homology) with the 13 known subtypes of feIFN-ω ( Figure 1d ). The sequence homology shared between the feIFN-ωa and feIFN-ωb genes was only 83.01% (Figure 1b) , indicating that the two genes identified in this study were novel feIFN-ω genes.