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

Inactive Publication Date: 2014-01-29
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

GCRV-HZ08 (type II) is a new isolate, and the functions and effects of its segments and their encoded proteins have not been studied in depth

Method used

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  • Recombinant protein coded by grass carp reovirus (GCRV) type-II S10 gene, polyclonal antibody prepared from recombinant protein and application of recombinant protein
  • Recombinant protein coded by grass carp reovirus (GCRV) type-II S10 gene, polyclonal antibody prepared from recombinant protein and application of recombinant protein
  • Recombinant protein coded by grass carp reovirus (GCRV) type-II S10 gene, polyclonal antibody prepared from recombinant protein and application of recombinant protein

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Embodiment

[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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Abstract

The invention discloses a recombinant protein coded by a grass carp reovirus (GCRV) type-II S10 gene, a polyclonal antibody prepared from the recombinant protein and an application of the recombinant protein. The amino acid sequence of the GCRV type-II S10 gene-coded protein is shown by SEQ ID No:2, and the nucleotide sequence coding the protein is shown by SEQ ID No:1. The recombinant protein disclosed by the invention has good immunogenicity; compared with the proteins of other structures of GCRV, the specific antibody valence generated by inducing an immune animal is higher. Further tests indicate that by immunizing the grass carp with the S10 recombinant protein, the grass carp can be induced to generate high specific antibody, and certain immune protection effect can be realized against the attack of a virulent strain of GCRV. Therefore, the study and application of the GCRV type-II S10 gene-coded protein are of vitally important significance to the development of a novel vaccine for the hemorrhagic disease of grass carp and an immunological detection kit, and a new effective solution is provided for the hemorrhagic disease of grass carp.

Description

Technical field [0001] The invention is a biological gene engineering field, which involves the reorganization S10 antigen protein coded by the grass carp virus Ⅱ S10 gene, and the polyxon antibody and its application prepared by the S10 antigen protein. Background technique [0002] The grass carp hemorrhage caused by the Grass Craw Reovirus (GCRV) has caused huge economic losses to the grass carp breeding industry in my country, which seriously restricts the healthy development of grass carp breeding in my country.GCRV has double -layer clothing shells, the average diameter of the virus particles is 60 nm to 70 nm, the twenty -steroid is symmetrical, no capsule film, and the genomic group consists of a double -chain RNA with a 11 -point segment.The genome is divided into three groups according to the size of the molecular weight, named L1-L3, M4-M6 and S7-S11, respectively.These 11 RNA segments can encode 12 polypeptide proteins, 7 of which are named VP1-VP7 respectively, the r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/14C07K16/10C12N15/46A61K39/15A61K39/42A61P31/14G01N33/68G01N33/569
CPCA61K39/15A61K2039/505C07K14/005C07K16/10C12N2720/12222C12N2720/12234G01N33/5091G01N33/56983G01N2333/14
Inventor 曾伟伟王庆吴淑勤王英英梁红茹石存斌李莹莹刘春
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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