System, method and application for saving swine enteric alphacoronavirus
A coronavirus, pig intestinal technology, applied in the system field of saving pig intestinal type A coronavirus, to achieve the effect of wide application prospects
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[0036] 1. Amplification of the target fragment of SADS-CoV / SeACoV full-length infectious clone
[0037] According to the gene sequence of porcine enteric alpha-coronavirus SeACoV CH / GD-01 / 2017 / P2 strain published on Genbank (GenBank accession no. MF370205, figure 1 ), the full length of the virus genome is 27155 nt, and the homology of the bat enteric A-coronavirus HKU2 found in the same area 10 years ago and infecting the Chinese horseshoe bat at the whole genome nucleic acid level is 94.9%. The cell infection supernatant sample of the cell stable passage strain SADS-CoV / SeACoV-P10, the Trizol method extracted the RNA and then reverse-transcribed to obtain cDNA, using 16 pairs of primers provided by the present invention (see Table 1 for the primer sequence), amplified to obtain the corresponding The 16 full-length genome fragments of SADS-CoV / SeACoV-P10 were cloned by pCR-Blunt vector, 3 single clone colonies were picked, plasmids were extracted, sequenced and spliced, and t...
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