RHVT (recombinant Herpesvirus of Turkey)-H9HA (H9 hemagglutinin) and construction method thereof

A technology of turkey herpes virus and construction method, which is applied in the field of recombinant virus vaccines, can solve the problems of HVT recombinant virus screening difficulties, animal resistance to drugs, interference with recombinant virus immune effects, etc.

Inactive Publication Date: 2015-10-28
YANGZHOU UNIV
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Problems solved by technology

Due to the strong cell-binding properties of HVT, this brings great difficulties to the screening of HVT recombinant viruses
At present, the most commonly used method is the marker gene insertion method to screen recombinant viruses. However, the presence of marker genes has brought many disadvantages, such as

Method used

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  • RHVT (recombinant Herpesvirus of Turkey)-H9HA (H9 hemagglutinin) and construction method thereof
  • RHVT (recombinant Herpesvirus of Turkey)-H9HA (H9 hemagglutinin) and construction method thereof
  • RHVT (recombinant Herpesvirus of Turkey)-H9HA (H9 hemagglutinin) and construction method thereof

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Experimental program
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Embodiment Construction

[0034] Step 1: Amplification of DNA Fragments

[0035] Biomaterial preparation:

[0036] HVT Fc-126 strain [Hu Chuanwei, Peng Daxin, Zhang Rukuan, Liu Xiufan. Immunological efficacy of recombinant fowlpox virus expressing chicken Marek's disease virus gB gene and turkey herpes virus bivalent freeze-dried vaccine[J]. Chinese Journal of Veterinary Medicine, 2006, 01:11‐13.], preserved by the Key Open Laboratory of Livestock and Poultry Infectious Diseases, Ministry of Agriculture, Yangzhou University.

[0037] The positive cloning plasmid pPWJM containing the HA gene of H9N2 subtype AIV epidemic strain WJ57 [Ding Pingyun, Liu Xiufan et al. Construction and rescue of H9 subtype avian influenza and Newcastle disease recombinant vaccine strains[J]. China Poultry, 2013, 16: 16-20.], constructed and preserved by the Laboratory of Livestock and Poultry Infectious Diseases, Ministry of Agriculture, Yangzhou University.

[0038] pEGFP-C1 eukaryotic expression plasmid: purchased from B...

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Abstract

The invention relates to an rHVT (recombinant Herpesvirus of Turkey)-H9HA (H9 hemagglutinin) for expressing H9 subtype AIV (avian influenza virus) HA protein and a construction method thereof. The collection number of a vaccine strain rHVT-H9HA is CGMCC No: 10907. A GFP (green fluorescent protein) expression cassette is separated from a carrier pEGFP-C1 (plasmid enhanced green fluorescent protein) and is inserted into an HVT genome, and a recombinant virus rHVT-GFP is obtained. Through homologous recombination, a GFP gene of the rHVT-GFP is replaced with an HA gene of H9N2 subtype AIV epidemic strain A/Chicken/Jiangsu/WJ57/2012, a recombinant virus without fluorescence is selected, and the rHVT-H9HA for stably expressing the H9 subtype AIV HA gene is obtained. The recombinant virus strain is low in cost, good in safety, long in immunity period and suitable for large-scale production of vaccine and can be used for making the vaccine.

Description

technical field [0001] The invention relates to the field of recombinant virus vaccines, more specifically, the invention relates to a turkey herpes virus strain rHVT-H9HA expressing the H9 subtype HA gene of avian influenza, which is used for the development of the vaccine. The invention also provides a method for developing the recombinant turkey herpes virus vaccine strain. Background technique [0002] Among the many pathogens that cause poultry diseases, avian influenza virus (Avian influenza virus, AIV) and Marek's disease virus (Marek's disease virus, MDV) occupy an important position. Prevention and control of outbreaks of avian influenza and Marek's disease in poultry is of great significance to animal health, public health and the world economy. [0003] Avian influenza (Avian influenza, AI) is a viral infectious disease that seriously endangers the development of the global poultry industry. AIV belongs to the genus of influenza virus A in the Orthomyxoviridae f...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/85C12R1/93
Inventor 刘秀梵郝小利吴艳涛王晓泉胡顺林胡娇顾敏
Owner YANGZHOU UNIV
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