Preparation method, application and preparation primer of influenza virus HA fusion peptide and helix A protein

A technology of influenza virus and fusion peptide, applied in the direction of virus peptide, fusion peptide, chemical instruments and methods, etc., to achieve good immunogenicity effect

Inactive Publication Date: 2018-12-25
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, whether the fusion peptide of HA and the helix A region can be used as a broad-spectrum subunit vaccine for influenza virus is still inconclusive

Method used

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  • Preparation method, application and preparation primer of influenza virus HA fusion peptide and helix A protein
  • Preparation method, application and preparation primer of influenza virus HA fusion peptide and helix A protein
  • Preparation method, application and preparation primer of influenza virus HA fusion peptide and helix A protein

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Embodiment

[0030]1. Design and amplify the primers containing the pGEX-6p-1 linearized vector, influenza virus HA fusion peptide and helix A gene fragment: the upstream primer for amplifying the pGEX-6p-1 linearized vector is located at positions 1022-1047 of the pGEX-6p-1 plasmid ; and with an extra TAA base at the 5' end; the downstream primer for amplifying the pGEX-6p-1 linearized vector is located at position 916-941 of the pGEX-6p-1 plasmid; and with an extra CAT base at the 5' end. Amplified influenza virus HA fusion peptide and helix A gene upstream primers added ATG GGA AGC as a strong promoter including 17 bases of HA fusion peptide and helix A gene, and 13 bases at the 5' end with pGEX-6p-1 Additional bases for the reverse complement of the downstream primers of the linearized vector; amplify the influenza virus HA fusion peptide and the helix A gene The downstream primers include the HA fusion peptide and the helix A gene stop codon TAA and its first 15 bases, and at the 5' en...

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Abstract

The invention discloses a preparation method, application and preparation primer of an influenza virus HA fusion peptide and helix A protein expression vector. The principle and key technique herein are to scientifically design and amplify a pGEX-6P-1 linear vector and influenza virus HA fusion peptide-helix A gene fragment; commercial recombinant enzyme Exnase TM II is subjected directly to in-vitro quick recombinant cloning of HA fusion peptide-helix A without enzyme-cut and link-up connection; Escherichia coli is converted and expressed under IPTG induction, and fusion expression of influenza virus HA fusion peptide-helix A protein with GST is achieved; the purified HA fusion protein-helix A fusion protein is attained.

Description

technical field [0001] The invention relates to a preparation method and application of influenza virus HA fusion peptide and helix A protein. The invention can directly provide HA fusion peptide and helix A fusion protein as immunogens to obtain anti-HA fusion peptide and helix A polyclonal antibody. The invention will lay a foundation and provide materials for further exploring whether the fusion peptide of HA and the helix-A region can be used as an effective epitope to stimulate the body to produce neutralizing antibodies, and to evaluate the broad-spectrum influenza neutralizing subunit vaccine. Background technique [0002] The use of vaccines and antiviral drugs has made positive contributions to the prevention and control of influenza viruses. However, due to the continuous variation of influenza virus antigens and the continuous emergence of recombinant viruses, human influenza pandemics and avian influenza outbreaks often occur, seriously affecting human health an...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/70C12N15/62C07K19/00
CPCC07K14/005C07K2319/00C12N15/70C12N2760/16122
Inventor 叶建强李占萍邵红霞秦爱建万志敏
Owner YANGZHOU UNIV
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