Recombinant fowl pos virus vaccine rFPV 1218AIH5/H9 and its construction process and use

A technology of rfpv-1218aih5 and fowlpox virus, applied in the field of recombinant fowlpox virus combined vaccine, can solve the problem of destroying IL-18 binding protein gene and so on

Inactive Publication Date: 2007-11-07
YANGZHOU UNIV
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Problems solved by technology

[0008] According to the previous research of our laboratory, we found that the non-essential region FPV7S of the insertion v

Method used

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  • Recombinant fowl pos virus vaccine rFPV 1218AIH5/H9 and its construction process and use
  • Recombinant fowl pos virus vaccine rFPV 1218AIH5/H9 and its construction process and use
  • Recombinant fowl pos virus vaccine rFPV 1218AIH5/H9 and its construction process and use

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Embodiment Construction

[0057] The following biological material Escherichia coli p12-18 was deposited in the General Microorganism Center of China Committee for the Collection of Microorganisms on June 6, 2005. The preservation number is: CGMCC NO: 1385, and its length is: 10450bp.

[0058] Step 1: Amplification, cloning and sequence analysis of H5 subtype AIV HA gene

[0059] According to the published sequence of AIV HA gene of H5 subtype, the primers for PCR amplification of H5A gene were designed. Primers were synthesized by Bao Biological Engineering (Dalian) Co., Ltd.

[0060] Amplification primers of the H5A gene: BamHI site and Kozak sequence were introduced into the 5' end of the upstream primer PH5A1, BamHI and HindIII sites were introduced into the 5' end of the downstream primer PH5A2, and the transcription termination signal of the early gene of the poxvirus genome was also introduced. The upstream and downstream primers span about 1.7kb, covering the entire coding frame of the H5A gen...

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Abstract

The present invention relates to recombinant fowl pos virus vaccine constructing technology, and is especially recombinant fowl pos virus vaccine rFPV-1218AIH5/H9 and its construction process and use. Through setting the H5 subtype-containing AIV HA gene segment and H9 subtype-containing AIV HA gene segment separately in the downstream of respective promoter, back-to-back connecting serially, inserting into fowl pos virus expressing vector p12-18 containing FPV copying non-essential segment FPV12-18 and reporter gene LacZ to constitute transfer vector; transfecting wt-FPV-infecting chick embryo fibroblast with the transfer vector; and homogeneous recombination, virus vaccine with co-expressed H5 and H9 subtype AIV HA gene and free from interference of maternal antibody is obtained.

Description

technical field [0001] The invention relates to a recombinant fowlpox virus vaccine, in particular to a recombinant fowlpox virus combined vaccine that co-expresses the main protective antigen genes of H5 and H9 subtype avian influenza viruses by using a new replication non-essential region of the fowlpox virus. Background technique [0002] Fowlpox Virus (Fowlpox Virus, abbreviated: FPV) is a member of the family Poxviridae and the genus Fowlpoxvirus, and is the largest virus among animal viruses discovered so far. The most unique property is that they replicate in the cytoplasm of infected cells and not in the nucleus. Mature virions are brick-shaped with a size of 250×350nm. The genome of FPV is a double-stranded linear DNA with a size of about 300kb and a molecular weight of about 2-4×10 5 KD, the G+C content reaches 35%, and its DNA is not infectious. Through recombinant DNA technology, more and more researchers have paid attention to the development of poultry geneti...

Claims

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Application Information

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IPC IPC(8): A61K39/275A61P31/16C12N15/85
Inventor 刘秀梵陈素娟孙蕾刘武杰
Owner YANGZHOU UNIV
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