Infusion protein of IDB (Infectious Bursal Disease) virus VP2 protein and LTB (Leukotrienes B) and application thereof

A fusion protein and virus technology, applied in the direction of antiviral agents, virus antigen components, medical preparations containing active ingredients, etc., can solve the problems that the protection rate is only 10%~70%, and cannot provide sufficient protection.

Active Publication Date: 2013-03-27
QINGDAO VLAND BIOTECH INC +1
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

In the past ten years, due to the emergence of super-virulent strains and variants of chicken infectious bursal disease virus IBDV, antigenic variation and diversity of serotypes, the current commercial vaccines cannot provide sufficient protection. Only 10%~70%

Method used

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  • Infusion protein of IDB (Infectious Bursal Disease) virus VP2 protein and LTB (Leukotrienes B) and application thereof
  • Infusion protein of IDB (Infectious Bursal Disease) virus VP2 protein and LTB (Leukotrienes B) and application thereof
  • Infusion protein of IDB (Infectious Bursal Disease) virus VP2 protein and LTB (Leukotrienes B) and application thereof

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Embodiment Construction

[0012] The main structural protein VP2 protein of IBDV is the main host protective antigen, and Escherichia coli heat-labile enterotoxin LTB is a good mucosal immune adjuvant. The present invention uses the overlap extension PCR method to connect the two genes through a linker and clone Transform BL21(DE3) onto the pET-28a vector, and the VP2-LTB fusion protein is expressed after being induced by IPTG. After the expression bacteria are homogenized under high pressure, the supernatant is centrifuged to obtain the VP2-LTB protein solution, and mineral oil is added. The adjuvant is made into an oil emulsion vaccine, which can be used to prevent chicken infectious bursal disease, and the effect is better than that of the VP2 subunit vaccine.

[0013] The fusion protein of the present invention will be described in detail below in conjunction with specific examples.

[0014] 1. Screening of VP2 gene

[0015] A pair of primers for amplifying chicken infectious bursal disease virus ...

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Abstract

The invention relates to an infusion protein of an IDB (Infectious Bursal Disease) virus VP2 protein and an LTB (Leukotrienes B) of enterotoxin of escherichia coli, wherein the amino acid sequence of the infusion protein is SEQ ID NO: 1. According to the invention, the supervirulent VP2 protein of the IBDV (Infectious Bursal Disease Virus) is expressed with the LTB protein of enterotoxin of escherichia coli with a mucosal immunologic adjuvant in a fusion manner, and an expression product is emulsified to prepare a vaccine to make SPF chickens which are 21 days old immune. An immune reference and a non-immune reference for a subunit vaccine of VP2 are set. After 7 days of immunity, the lymphocyte transformation rate is determined, and after 21 days of immunity, blood is collected to determine the valence of antibody of immunodiffusion of serum agar and toxic substances are counteracted together with the non-immune reference. The result shows that vaccine immunity groups can resist virulent attach. The lymphocyte transformation rate and the serum valence of antibody are remarkably higher than the immune reference group of VP2, so that the infusion protein has an excellent immunogenicity.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering subunit vaccines, and specifically relates to a fusion protein of IBD virus VP2 protein and LTB and application thereof, that is, a fusion protein of chicken infectious bursal disease virus VP2 protein and Escherichia coli enterotoxin LTB and application thereof. Background technique [0002] Chicken infectious bursal disease (Infectious Bursal Disease, IBD) mainly invades the central immune organ of chickens, the bursa of Fabricius, which leads to immunosuppression, increases the susceptibility of the body to other pathogens, and reduces the reactivity to other vaccines. Foreign scholars The disease is vividly called "AIDS" of chickens. Although scholars from various countries have made great achievements in the research of IBD vaccines, none of them can fundamentally reverse the current situation of the increasingly serious IBD epidemic. In the past ten years, due to the emergence o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62A61K39/12A61K39/39A61P31/14
Inventor 王雷凌红丽王宏华王睿智
Owner QINGDAO VLAND BIOTECH INC
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