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Molecular design of IBDV (Infectious Bursal Disease Virus) antigen epitope and HBcAg (Hepatitis B Core Antigen) chimeric gene

A chimeric gene, antigenic epitope technology, applied in the direction of viral antigenic components, recombinant DNA technology, microorganism-based methods, etc., can solve the problem of small molecular weight of multi-epitopes, expression, immunogenicity and immune efficacy need to be further improved, etc. problem, to achieve the effect of improving stability

Inactive Publication Date: 2010-09-01
中国人民解放军南京军区军事医学研究所 +1
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Problems solved by technology

However, the molecular weight of the multi-epitope obtained by simply using the method of cascading antigenic epitopes is small, and its expression, immunogenicity and immune efficacy need to be further improved.

Method used

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  • Molecular design of IBDV (Infectious Bursal Disease Virus) antigen epitope and HBcAg (Hepatitis B Core Antigen) chimeric gene
  • Molecular design of IBDV (Infectious Bursal Disease Virus) antigen epitope and HBcAg (Hepatitis B Core Antigen) chimeric gene
  • Molecular design of IBDV (Infectious Bursal Disease Virus) antigen epitope and HBcAg (Hepatitis B Core Antigen) chimeric gene

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Embodiment Construction

[0039] 1 Experimental materials

[0040] The recombinant prokaryotic expression plasmid pET-5epis containing the mimic IBDV multi-epitope gene 5epis was constructed by our laboratory [known and public, Wang YS (王永山), et al (etc.). Development of a multi-mimotope peptide as avaccine immunogen for infectious bursal disease virus. Vaccine, 2007, 25(22): 4447-4455.].

[0041] The chicken anti-IBDV high immune serum is obtained by using IBD attenuated vaccine (B87 strain, purchased from Nanjing Tianbang Biotechnology Co., Ltd.) to immunize SPF chickens (purchased from Nanjing Tianbang Biotechnology Co., Ltd.) for 5 times, using DEAE cellulose (DE32 ) Purification (Liu Yubin, Editor-in-Chief Gou Shijin. Experimental Technology of Animal Immunology. Changchun: Jilin Science and Technology Press, 1989, 8-15);

[0042] Preparation of rabbit anti-chicken IgG antibody and HRP-labeled rabbit anti-chicken IgG antibody reference literature (Liu Yubin, Gou Shijin editor. Experimental technology of...

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Abstract

The invention relates to a molecular design of an IBDV (Infectious Bursal Disease Virus) antigen epitope and HBcAg (Hepatitis B Core Antigen) chimeric gene, belonging to the field of biological pharmacy. The molecular design comprises the following steps of: establishing an expression plasmid pET-mHBc of an HBcAg-modified gene by applying an overlap extension PCR (Polymerase Chain Reaction) technology; directionally inserting an infectious bursal disease virus gene 5epis into the expression plasmid mHBC gene, acquiring an expression plasmid pEt-mHBc-5epis, and expressing the pEt-mHBc-5epis in colibacillus, wherein the molecular weight of recombinant chimeric protein is about 29kDa, and the expression level accounts for about 47.6 percent of total bacterial protein, the diameters of virus-like particles are about 60 to 80nm, and the antigen valence reaches 2.0*102. The 5epis and HBcAg chimeric gene which is successfully established by adopting the molecular design efficiently expresses the recombinant chimeric protein with the IBDV antigen reactivity in the colibacillus, forms the virus-like particles, and has the application potential of becoming a granulated epitope-based IBD (Infectious Bursal Disease) genetic engineering vaccine.

Description

1. Technical Field [0001] The present invention relates to the molecular design of IBDV epitope and HBcAg chimeric gene, in particular to the molecular design and technical route of infectious bursal disease virus (IBDV) epitope and hepatitis B virus core antigen (HBcAg) chimeric gene , Gene sequences, expression plasmids and their expression products belong to the field of biopharmaceuticals. 2. Background technology [0002] Infectious Bursal Disease (IBD) is one of the most serious poultry diseases today. It is caused by Infectious Bursal Disease Virus (IBDV) to damage the central immune organs of chicks. The bursa is the main feature. The disease not only causes the death of diseased animals, but also causes the body's immune suppression, which reduces the body's immune defense ability and fails the vaccination. The annual economic loss caused by the onset of IBD, immune failure, and immunosuppression is huge [Wang Yongshan, etc.. Molecular characteristics of the VP2 gene o...

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Application Information

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IPC IPC(8): C12N15/62C07K19/00C12N15/70C12P21/02G01N33/569A61K39/12A61P31/14C12R1/19
Inventor 王忠灿王永山刘小娟欧阳伟王晓丽诸玉梅唐雨德
Owner 中国人民解放军南京军区军事医学研究所
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