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Recombined chicken pox virus vaccine rFPV-12LSH5H9, its construction method and use

A fowlpox virus, rfpv-12lsh5h9 technology, applied in the field of recombinant fowlpox virus combined vaccine, can solve the problem of destroying IL-18 binding protein gene

Inactive Publication Date: 2006-02-22
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 vector p11S constructed by our laboratory is just in the open reading frame, which may destroy the IL-18 binding protein gene

Method used

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  • Recombined chicken pox virus vaccine rFPV-12LSH5H9, its construction method and use
  • Recombined chicken pox virus vaccine rFPV-12LSH5H9, its construction method and use
  • Recombined chicken pox virus vaccine rFPV-12LSH5H9, its construction method and use

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Embodiment Construction

[0057] The following will be described in detail in conjunction with the implementation examples and the accompanying drawings, but the following implementation examples are to further explain rather than limit the present invention.

[0058] The fowlpox virus expression vector p12-18 involved below was deposited in the General Microorganism Center of China Committee for Culture Collection of Microorganisms on June 6, 2005. The preservation number is: CGMCC NO: 1385, and its length is: 10450bp.

[0059] Step 1: Amplification, cloning and sequence analysis of H5 subtype AIV HA gene

[0060] 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.

[0061] Amplification primers of the H5A gene: the 5' end of the upstream primer PH5A1 introduces the BamHI site and Kozak sequence (as shown in the box), the 5' end of the downstream primer ...

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Abstract

Disclosed is a recombined chicken pox virus vaccine rFPV-12LSH5H9, its construction method and use, wherein the construction process consists of placing AIV HA gene fragment containing H5 hypotype and H9 hypotype onto the downstream of right promotors, inserting into chicken poxviridae expression vector p12-18 containing FPV duplicated non-indispensable fragment FPV1-2 and reported genes through backside cascade connection to construct transition vector p12LSH5H9, then transforming chick embryo desmocyte infected with wt-FPV with the transition vector p12LSH5H9, and obtaining co-expression H5 and H9 hypotype AIV HA gene rFPV-12LSH5H9 through consanguinity recombination.

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): A61K39/275C12N15/09C12N15/86A61P31/16
Inventor 刘秀梵陈素娟孙蕾刘武杰
Owner YANGZHOU UNIV
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