Recombinant turkey herpesvirus and application thereof

A turkey herpes virus and virus technology, applied in the field of animal virology, can solve the problems of poor immune protection and damage to the bursa of Fabricius in chickens with maternal antibodies

Active Publication Date: 2012-08-15
ZHAOQING DAHUANONG BIOLOGIC PHARMA +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The attenuated live vaccine is safe for SPF chickens, but the immune protection for chickens with maternal antibodies is poor
Although the immune protection of moderately virulent and highly virulent vaccines is much higher, they will damage the bursa of Fabricius due to their certain pathogenicity.
At the same time, due to the interference of maternal antibodies and the characteristics of IBDV, traditional vaccines can form an inevitable immune blank period in practical applications.

Method used

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  • Recombinant turkey herpesvirus and application thereof
  • Recombinant turkey herpesvirus and application thereof
  • Recombinant turkey herpesvirus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1 The preparation of the expression cassette containing the VP2 (IBDV-VP2) gene of chicken infectious bursal virus:

[0045] The IBDV-VP2 gene expression cassette sequence was synthesized by Nanjing GenScript Biotechnology Co., Ltd. The IBDV-VP2 expression cassette includes a promoter sequence (Murine cytomegalovirus enhancer DNA sequence), a chicken infectious bursal virus VP2 gene and an SV40 termination sequence. The promoter sequence (Murine cytomegalovirus enhancer DNA sequence) (ACCESSION NO.L06570) was designed according to the reference (K Dorsch-Hasler, A long and complex enhancer activates transcription of the gene coding for the highly abundant immediate early mRNA in murine cytomegalovirus, PNAS: 1985 vol.82 no.248325-8329; M Messerle, Structure and expression of murine cytomegalovirus immediate-early gene 2, J.Virol.March 1991 vol.65 no.31638-1643); IBDV VP2 sequence reference Cu-1wt strain design (ACCESSION NO.AF362747). The gene sequence of t...

Embodiment 2

[0046] The construction of embodiment 2 recombinant virus rHVT-VP2:

[0047] Utilize the method for homologous recombination to construct recombinant virus rHVT-VP2, the steps are as follows:

[0048] (1) On the basis of the above-mentioned plasmid PUC19-VP2, the inventor constructed a recombinant plasmid PUC19-UL-VP2 containing a non-essential region of the HVT genome as a homologous plasmid, and its structure is shown in the attached Figure 4 As shown, the specific steps are as follows: Except for the following content, other construction methods in the prior art are adopted,

[0049] Extract the HVT virus genomic DNA according to conventional methods, and design and amplify the left and right homology arms of the UL55 and LORF4 regions according to the complete gene sequence of the HVT-FC126 strain published in Genebank. The primers used to amplify the left homology arms are as follows: ULL- F: GTCGACacaagtaaaatacccaacac (a Sal I restriction site is added to the front end...

Embodiment 3

[0055] In vitro and in vivo stability experiments of embodiment 3 rHVT-VP2

[0056] Passage rHVT-VP2 in CEF cells for 50 times, and use the above primers (rHVT-VP2-JC-F; rHVT-VP2-JC-R) to amplify, the result can still amplify a 3083bp band, as shown in the attached Image 6 As shown, it shows that the invented rHVT-VP2 remains stable after 50 passages in cells.

[0057] SPF chickens were inoculated subcutaneously with 2500 PFU of rHVT-VP2. After 3, 4, 5 and 6 weeks of inoculation, peripheral blood was collected from the vaccinated chickens and inoculated with CEF cells, cultured for 5-7 days, and stained with anti-IBDV-VP2 monoclonal antibody after plaques grew. All plaques could express IBDV VP2 gene, which indicated that rHVT-VP2 was stable in vivo.

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Abstract

The invention relates to the field of animal virology, and provides a recombinant turkey herpesvirus (rHVT-VP2), which is actually a brand-new recombinant turkey herpesvirus rHVT-VP2 obtained by combining an expression box of an avian infectious bursal disease virus VP2 gene and a turkey herpesvirus HVT. The recombinant turkey herpesvirus is also a recombinant vaccine of Marek's disease and avianinfectious bursal disease. By inserting the VP2 gene into a gene complex of the HVT under control of promoter, the rHVT-VP2 not only has fine immune protection for avian Marek's disease, but also haspersistent protective immunity of induced resistance of IBDV (infectious bursal disease virus) for commercial chickens with high IBDV material antibody level.

Description

technical field [0001] The invention relates to the field of animal virology. The invention provides a recombinant turkey herpes virus (rHVT-VP2), whose genome contains chicken infectious bursal virus (IBDV) VP2 gene under the control of a promoter. Background technique [0002] Chicken Marek's disease (MD) and chicken infectious bursal disease (IBD) are two of the most common immunosuppressive diseases affecting poultry production. Among them, IBD is caused by chicken infectious bursal disease virus (Infectious bursal disease virus, IBDV), and it is an acute and highly contagious infectious disease that mainly harms young chickens aged 3 to 12 weeks. IBDV mainly causes damage to the bursa, the main immune organ of chickens, which leads to severe immunosuppression in chickens, increases the susceptibility of infected chickens to other diseases and reduces the immune response to vaccines, causing huge losses to the poultry industry. Loss (Mundt E, Beyer J, Müller H. Identifi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/40A61K39/295A61K39/245A61K39/12A61P31/14A61P31/22C12R1/93
Inventor 陈瑞爱
Owner ZHAOQING DAHUANONG BIOLOGIC PHARMA
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