Porcine deltacoronavirus strain and application thereof
A technology of coronavirus and strains, applied in the field of porcine delta coronavirus strains, can solve the problems of undeveloped effective vaccines or drugs, and achieve the effect of strong toxicity and good immune effect
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[0034] 1 Disease material and cells
[0035] The disease materials were collected from Xinjiang, China, and 12 samples containing PDCoV-positive strains were used for the isolation and identification of PDCoV; LLC-PK-1 cells were cryopreserved by our laboratory. The cells were subcultured in MEM medium containing 10% FBS and 1% double antibody.
[0036] 2 Determination of S genome sequence of PDCoV CH / XJYN03 / 2016 strain
[0037] According to the published PDCoV sequence comparison results, the conserved regions were selected, and the corresponding amplification primers were designed to amplify the S genome sequence of PDCoV. The primers are shown in Table 1. After the CH / XJYN03 / 2016 strain field feces samples and cell culture virus liquid RNA were extracted, RT-PCR reactions were performed using S genome-specific primers. The pCR-XL-2TOPO vector for blunt end connection was selected for subcloning of the PCR fragment, and the correct plasmid identified by enzyme digestion wa...
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