Recombinant protein coded by grass carp reovirus (GCRV) type-II S10 gene, polyclonal antibody prepared from recombinant protein and application of recombinant protein
A technology of reovirus and polyclonal antibody, which is applied in the direction of antiviral immunoglobulin, virus antigen components, virus/phage, etc., can solve the problem that the function and function of the encoded protein have not been studied in depth, and overcome the cumbersome operation process , good immune protection effect, good immunogenicity effect
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[0028] 1. RNA extraction and S10 gene fragment amplification of grass carp reovirus HZ08 strain
[0029]According to the instructions of the Qiagen RNA extraction kit (purchased from Qiagen, product number: 74104), RNA was extracted from CIK cells infected with GCRV HZ08 strain virus, and reacted with TAKARA (purchased from Dalian Bao Biological Engineering Co., Ltd., product number: ). The transcription kit reverse transcribes viral RNA into cDNA. The GCRV HZ08 virus strain was preserved by the Fish Disease Department of the Pearl River Fisheries Research Institute.
[0030] Design a pair of primers according to the S10 gene sequence of GCRV-HZ08 strain in GenBank (GenBank: GU350747.1), upstream primer: 5'-ATA GGA TCC ATG GCG GGT GTG TCT CT CAA CA - 3' (SEQ ID NO: 3); downstream primer: 5'-GCT AAG CTT CAG CAT CTG CGC AAA TAT ACG TC -3' (SEQ ID NO: 4), insert BanHI and HandIII restriction sites (underlined genes) into the upstream and downstream primer sequences respec...
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