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Recombinant herpesvirus turkey live vector vaccine capable of simultaneously expressing classical strain and variant infectious bursal disease virus VP2 protein

A technology of turkey herpes virus and classic strains, applied in the direction of virus/bacteriophage, virus, vaccine, etc., can solve problems such as weight loss, interference with immune effect, and increased feed-to-meat ratio, so as to improve antibody levels, ensure immune effects, and improve The effect of uniformity

Active Publication Date: 2022-03-01
扬州优邦生物药品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the lethality of the new mutant strain of IBDV is low, the irreversible bursal damage and severe immunosuppression caused by the IBDV lead to low immunity of the chickens, which increases the chance of infection of other pathogens, affects the growth of the chickens, and leads to weight loss , poor uniformity, increased feed-to-meat ratio, etc., seriously affecting economic benefits
On the other hand, new variants of IBDV will interfere with the immune effect of other important disease vaccines

Method used

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  • Recombinant herpesvirus turkey live vector vaccine capable of simultaneously expressing classical strain and variant infectious bursal disease virus VP2 protein
  • Recombinant herpesvirus turkey live vector vaccine capable of simultaneously expressing classical strain and variant infectious bursal disease virus VP2 protein
  • Recombinant herpesvirus turkey live vector vaccine capable of simultaneously expressing classical strain and variant infectious bursal disease virus VP2 protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1: Determination of Infectious Bursal Virus VP2 Gene

[0068] The VP2 gene of the classic strain of infectious bursal virus is obtained from the relevant sequence of the subunit vaccine produced by our company, through the online molecular biology software of GenScript, fused with LTB by molecular software, and then optimized with chicken source as the host A nucleotide sequence and named LTB-VP2C. The VP2 protein gene sequence of the infectious bursal virus mutant strain was isolated by the technical service department of our company, sequenced and compared by NCBI, and a nucleotide sequence was optimized by using the same online codon optimization tool with chicken source as the host , named LTB-VP2V.

Embodiment 2

[0069] Embodiment 2: Preparation of recombinant virus rHVT-US2-VP2C

[0070] 1. Genome extraction of turkey herpes virus: extract the total DNA of turkey herpes virus by traditional DNA extraction method (NCBI accession number: AF291866.1);

[0071] 2. Design and synthesis of homologous recombination homology arm transfer vector: According to the GenBank published turkey herpes virus US2 gene sequence, the left and right homology arms were designed with 1200 bp each, and the pec-EGFP expression cassette was introduced in the middle of the homology arms (by pec Promoter, EGFP gene and US2 self-terminator) and pec-VP2C (composed of pec promoter, LTB-VP2C gene and US2 self-terminator), synthesized into pBluescriptⅡKS by Nanjing GenScript Biotechnology Co., Ltd. The recombinant plasmids were named pB-LR-US2-EGFP and pB-LR-US2-LTB-VP2C;

[0072] 3. Rescue of recombinant turkey herpes virus: Co-transfect CEF cells with the transfer vector pB-LR-US2-EGFP and the turkey herpes virus ...

Embodiment 3

[0080] Example 3: Preparation of recombinant virus rHVT-US2-US10-LTB-VP2C-VP2V

[0081] 1. Genome extraction of turkey herpes virus: Extract the total DNA of the recombinant virus rHVT-US2-LTB-VP2C in Example 2 using traditional DNA extraction methods;

[0082]2. Design and synthesis of homologous recombination homologous arm transfer vector: According to the GenBank published turkey herpes virus US10 gene sequence, the left and right homologous arms were designed with 1200 bp each, and the pec-EGFP expression cassette was introduced in the middle of the homologous arms (by pec promoter, EGFP gene and US10 self-terminator) and pec-LTB-VP2V (composed of pec promoter, LTB-VP2V gene and US10 self-terminator), synthesized into pBluescriptⅡKS by Nanjing GenScript II KS. The recombinant plasmids were named pB-LR-US10-EGFP and pB-LR-US10-LTB-VP2V;

[0083] 3. Rescue of recombinant turkey herpes virus: co-transfect CEF cells with the transfer vector pB-LR-US10-EGFP and the turkey her...

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Abstract

The invention discloses a recombinant herpesvirus turkey live vector vaccine capable of simultaneously expressing classical strain and variant strain infectious bursal disease virus VP2 protein, and belongs to the field of veterinary biological products. According to the invention, a VP2 protein and LTB fusion expression cassette containing an infectious bursal disease virus classical strain is knocked into a US2 virus replication non-essential region of a turkey herpesvirus, and a VP2 protein and LTB fusion expression cassette of a variant strain is inserted into a US10 virus replication non-essential region. The finally obtained recombinant virus simultaneously and efficiently expresses LTB-VP2 fusion antigen protein of the IBDV classic strain and the variant strain in CEF cells. The vaccine prepared by the invention can improve the antibody level after immunization, improve the uniformity of the antibody after immunization and ensure the immunization effect of the vaccine, has the advantages of high efficiency, good safety and lifelong immunization after one-time inoculation, does not cause clinical reaction and pathological damage to chicks, and has the vaccine protection rate of 100%.

Description

technical field [0001] The invention relates to a recombinant turkey herpes virus live vector vaccine that simultaneously expresses the VP2 protein of a classic strain and a mutant strain of infectious bursal virus, belonging to the field of veterinary biological products. Background technique [0002] Infectious Bursal Disease Virus (IBDV) mainly infects chicks aged 3-6 weeks. After the infection, the immune organs of the chickens are severely damaged, causing the body to produce immunosuppression, and eventually lead to the death of the chicks. The high mortality rate caused by Infectious Bursal Disease (IBD) has caused serious economic losses to the global poultry industry. It is listed as the three major infectious diseases that endanger poultry health along with Newcastle disease and Marek's disease. [0003] There are two serotypes of IBDV, and serotype I is the main cause of IBD. Serum type I IBDV can be divided into four branches: classic strain, super strain, atten...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/63C12N15/40A61K39/245A61P31/14
CPCC12N7/00C07K14/005A61K39/12A61P31/14C12N2710/16021C12N2720/10022C12N2720/10034A61K2039/552Y02A50/30
Inventor 叶正琴丁国伟范娟李甜甜徐萍李群潘晨桑建君包菲何存亚魏荣荣李琛钱钟杨振董昌海李玉和潘杰宋庆庆
Owner 扬州优邦生物药品有限公司
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