Genetic and phylogenetic analysis
The nucleotide sequences of MuKV identified in the present study were compared to the corresponding sequences of other kobuvirus strains previously reported in the Genbank database using Basic Local Alignment Search Tool (BLAST;https://www.ncbi.nlm.nih.gov). Multiple sequences were aligned with the corresponding regions of kobuvirus reference strains retrieved from Genbank database using the CLUSTAL W program in Molecular Evolutionary Genetics Analysis (MEGA version 7.0, Oxford Molecular Ltd., UK). The pairwise nucleotide (nt) and amino acid (aa) identities among all sequences were calculated using MegAlign program (DNASTAR, Inc. USA, Wisconsin, Madison). The open reading frame (ORF) was predicted for the obtained genome sequences by ORF finder (https://www.ncbi.nlm.nih.gov/orffinder). Similarity plot analysis of the full-length polyprotein genome was performed using SimPlot 3.5.1 software. The RNA secondary structure prediction for 5’ and 3’ UTR was conducted by Mfold program (Zuker, 2003). Phylogenetic analyses were generated by maximum-likelihood method with 1000 bootstrap replicates using MEGA v7.0 and further visualized in FigTree v1.4.0 software. The model selections were based on the results of ’Find best DNA/Protein models’ in MEGA v7.0.