Recombined chicken pox virus vaccine rPFV-12LSH9A, preparation process and use thereof

A fowl pox virus, rfpv-12lsh9a technology, applied in recombinant DNA technology, antiviral agents, pharmaceutical formulations and other directions, can solve the problem of destroying the IL-18 binding protein gene and other problems, and achieve the effect of less side effects and increased practicability

CN1736481AInactive Publication Date: 2006-02-22YANGZHOU UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2006-02-22
Estimated Expiration
Not applicable · inactive patent

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Abstract

Disclosed is a recombined chicken pox virus vaccine rPFV-12LSH9A, preparation process and use, wherein the construction process consists of placing AIV HA gene fragment containing 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 to construct transition vector, then transforming chick embryo desmocyte infected with wt-FPV with the transition vector, and obtaining rFPV expressing H9 hypotype AIV HA gene through consanguinity recombination, i.e. recombinant chicken poxviridae vaccine rFPV-12LSH9A.
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Description

technical field

[0001] The invention relates to a recombinant fowlpox virus vaccine, in particular to a recombinant fowlpox virus vaccine for expressing H9 subtype avian influenza virus hemagglutinin gene by using a newly identified replication non-essential region of 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 genetically engineered recombinant l...

Examples

Embodiment Construction

[0041] 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.

[0042] This example relates to the fowlpox virus expression vector p12-18, which was deposited in the General Microbiology Center of China Microbiological Culture Collection Management Committee on June 6, 2005. The preservation number is: CGMCC NO: 1385, and its length is: 10450bp.

[0043] Step 1: Amplification, cloning and sequence analysis of H9 subtype AIV HA gene

[0044] According to the known sequence of H9 subtype AIV HA in our laboratory, PCR amplification primers of H9N2 subtype AIV HA gene were designed. Primers were synthesized by Bao Biological Engineering (Dalian) Co., Ltd.

[0045] Amplification primers of the H9A gene: the 5' end of the upstream primer PH9A1 introduces the BamHI site and Kozak sequence (as shown in the box); th...