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Recombination Newcastle vaccine strain rAI4-S1 for expressing infectious bronchitis virus S1 protein and generating method thereof

A technology for bronchitis and construction methods, applied in the direction of recombinant DNA technology, microorganism-based methods, viruses/bacteriophages, etc., can solve the problems of meat quality decline, inactivated vaccines that cannot provide protection, adverse reactions, etc.

Inactive Publication Date: 2013-12-25
YANGZHOU UNIV
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Problems solved by technology

However, inactivated vaccines cannot provide sufficient protection, and oil-emulsion adjuvants can cause local adverse reactions, resulting in decreased meat quality and animal discomfort

Method used

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  • Recombination Newcastle vaccine strain rAI4-S1 for expressing infectious bronchitis virus S1 protein and generating method thereof
  • Recombination Newcastle vaccine strain rAI4-S1 for expressing infectious bronchitis virus S1 protein and generating method thereof
  • Recombination Newcastle vaccine strain rAI4-S1 for expressing infectious bronchitis virus S1 protein and generating method thereof

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Embodiment Construction

[0028] The recombinant Newcastle disease vaccine strain rAI4-S1 expressing infectious bronchitis virus S1 protein of the present invention was preserved on August 5, 2013 in the General Microbiology Center of China Microbiological Culture Collection Management Committee (Address: No. 1 Beichen West Road, Chaoyang District, Beijing No. 3, Institute of Microbiology, Chinese Academy of Sciences), classified as Newcastle disease virus rAI4-S1, preservation number CGMCC No: 8054.

[0029] Construction step 1: Construction of a full-length expression clone expressing infectious bronchitis virus S1 protein using Newcastle disease virus AI4 as a vector

[0030] Biomaterial preparation:

[0031] Full-length expression clone pNDV / AI4 of Newcastle disease virus AI4 strain (Hu Z, Hu S, Meng C, et al. Generation of a Genotype VII Newcastle Disease Virus Vaccine Candidate with High Yield in Embryonated Chicken Eggs. Avian Diseases2011, 55(3) :1759-1769) was constructed and preserved by the...

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Abstract

The invention discloses a recombination Newcastle vaccine strain rAI4-S1 for expressing infectious bronchitis virus S1 protein and a generating method of the recombination Newcastle vaccine strain rAI4-S1. The conservation number of the recombination Newcastle vaccine strain rAI4-S1 is CGMCC No: 8054. According to the generating method of the recombination Newcastle vaccine strain rAI4-S1, a built reverse genetic manipulation platform of a gene VII type NDV attenuated virus AI4 strain is utilized, the sequence revealed in the SEQ ID NO.7 is inserted into an AI4 strain genome full-length transcription carrier pNDV / AI4, and then the recombination Newcastle vaccine virus genome full-length cDNA clonal pNDV / AI4-S1 containing infectious bronchitis virus S1 genes is obtained. According to a recombination virus obtained through transfection, a chick embryo has high breeding titer, the S1 protein still can be expressed after continuous passage, and the generating method is suitable for large-scale production of vaccines and can be used for processing the vaccines.

Description

technical field [0001] The invention relates to the application of reverse genetic technology to rescue a recombinant vaccine strain rAI4-S1 expressing infectious bronchitis virus S1 protein with Newcastle disease virus as a carrier, and to use it for vaccine development. Background technique [0002] Reverse genetics manipulation technique (Reverse genetics manipulation technique): For RNA viruses, reverse genetics technology refers to the construction of infectious molecular clones of RNA viruses in vitro, and the reverse transcription of viral genomic RNA into cDNA, at the DNA molecular level It is a research technique for assembling new RNA viruses from viral genome cDNA and various auxiliary proteins through various in vitro manual operations, also known as full-length infectious cDNA cloning technology, and is often referred to as "virus rescue". [Neumann G,Whitt M A,Kawaoka Y,et al.(2002).A decade after the generation of a negative-sense RNA virus from cloned cDNA–wha...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/50C12N15/63C12R1/93
Inventor 刘秀梵陈坚屠颉王晓泉胡顺林
Owner YANGZHOU UNIV
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