A thermostable avirulent Newcastle disease virus (NDV) strain TS09-C was developed by serial passage of V4 strain in BHK-21 cells. The complete genome sequence of strain TS09-C was determined and compared with the sequences of other NDV isolates representing different thermostable phenotypes. The TS09-C genome was 15,186 nucleotides long and consisted of eight genes in the order of 3'-NP-P/V/W-M-F-HN-L-5'. High levels of nucleotide and amino acid sequence identity existed among the thermostable NDV isolates. Thermostable strains TS09-C, V4, I-2, and Ulster all belonged to genotype I. The F protein cleavage site of strain TS09-C was (112)G-K-Q-R-R-L(117), with an isolated basic amino acid and a pair of contiguous basic amino acids, differing from that of its parental V4 strain ((112)G-K-Q-G-R-L(117)). Our characterization of the complete genome of thermostable avirulent NDV strain TS09-C may facilitate the development of a thermostable NDV reverse genetic system and further study of the mechanism of thermostability of NDV.
The variant virulent porcine epidemic diarrhea virus (PEDV) strain (YN15) can cause severe porcine epidemic diarrhea (PED); however, the attenuated vaccine-like PEDV strain (YN144) can induce immunity in piglets. To investigate the differences in pathogenesis and epigenetic mechanisms between the two strains, differential expression and correlation analyses of the microRNA (miRNA) and mRNA in swine testicular (ST) cells infected with YN15, YN144, and mock were performed on three comparison groups (YN15 vs Control, YN144 vs Control, and YN15 vs YN144). The mRNA and miRNA expression profiles were obtained using next-generation sequencing (NGS), and the differentially expressed (DE) (p-value < 0.05) mRNA and miRNA were obtained using DESeq R package. mRNAs targeted by DE miRNAs were predicted using the miRanda algortithm. 8039, 8631 and 3310 DE mRNAs, and 36, 36, and 22 DE miRNAs were identified in the three comparison groups, respectively. 14,140, 15,367 and 3771 DE miRNA–mRNA (targeted by DE miRNAs) interaction pairs with negatively correlated expression patterns were identified, and interaction networks were constructed using Cytoscape. Six DE miRNAs and six DE mRNAs were randomly selected to verify the sequencing data by real-time relative quantitative reverse transcription polymerase chain reaction (qRT-PCR). Based on bioinformatics analysis, we discovered the differences were mostly involved in host immune responses and viral pathogenicity, including NF-κB signaling pathway and bacterial invasion of epithelial cells, etc. This is the first comprehensive comparison of DE miRNA–mRNA pairs in YN15 and YN144 infection in vitro, which could provide novel strategies for the prevention and control of PED.
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