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S1 and N double-antigen protein recombinant plasmid capable of expressing IBV (Infectious Bronchitis Virus) and construction method and application thereof

A protein recombination, double antigen technology, applied in the field of animal medicine bioengineering research, can solve problems such as unreported

Active Publication Date: 2014-03-12
GUANGXI UNIV
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  • Abstract
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Problems solved by technology

At present, there are few reports on ubiquitin-enhanced immune response to poultry pathogens, especially for viral antigens.

Method used

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  • S1 and N double-antigen protein recombinant plasmid capable of expressing IBV (Infectious Bronchitis Virus) and construction method and application thereof
  • S1 and N double-antigen protein recombinant plasmid capable of expressing IBV (Infectious Bronchitis Virus) and construction method and application thereof
  • S1 and N double-antigen protein recombinant plasmid capable of expressing IBV (Infectious Bronchitis Virus) and construction method and application thereof

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Embodiment Construction

[0016] 1.1 Virus strains

[0017] IBV strain GX-YL5 (GenBank NO.HQ848267.1).

[0018] 1.2 Method

[0019] 1.2.1 Design of construction plasmid primers

[0020] Primers were designed with reference to the known sequences of GX-YL5 whole gene and chicken ubiquitin gene in GenBank. Different primers had enzyme cutting sites for EocR I, Apa I, Hind III, BamH I and Xho I respectively. Enzymes were purchased from Bao Biological Engineering (Dalian) Co., Ltd. The sequences of primers for amplification of Ub, N and S1 genes are listed in Table 1.

[0021] Table 1 Primer Sequence

[0022]

[0023] In Table 1, the part in italics is the restriction site, and the part in brackets is the linker connecting the two genes.

[0024] During the construction of the plasmid pVAX1-Ub-linker-N-S1, the stop codon "TGA" at the end of the IBV N gene was artificially deleted, and the stop codon "TAA" was artificially added at the end of the IBV S1 gene to make it normal Express.

[0025] 1.2...

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Abstract

The invention discloses an S1 and N double-antigen protein recombinant plasmid capable of expressing IBV (Infectious Bronchitis Virus) and a construction method and an application thereof. The inventor designs primers according to S1, N and Ub gene sequences of infectious bronchitis virus registered in GenBank, amplifies the S1 and N genes of China isolate GX-YL5 strain of the infectious bronchitis virus and a chicken Ub gene through PCR (Polymerase Chain Reaction), inserts the N and S1 genes of the China isolate GX-YL5 strain of the infectious bronchitis virus and the chicken Ub gene into an eukaryotic expression vector pVAX1 through a molecular biological method, thus constructing an expression plasmid capable of co-expressing the N and S1 genes and the chicken Ub gene and researches DNA vaccine Pvax1-Ub-linker-N-S1. Experiments prove that the vaccine has a significant curative effect of preventing IBV. The S1 and N double-antigen protein recombinant plasmid disclosed by the invention can be used for simultaneously inserting the S1 and N genes of IBV and the chicken Ub gene which are connected in series into the pVAX1 vector to prevent the infectious bronchitis virus.

Description

technical field [0001] The invention belongs to the field of animal medicine bioengineering research, and in particular relates to a recombinant plasmid expressing S1 and N double antigenic proteins of IBV and its construction method and application. Background technique [0002] Infectious bronchitis (IB) is one of the second-class poultry infectious diseases stipulated by the International Veterinary Bureau and my country. It is currently one of the most widely distributed and most common diseases. The disease is an acute, highly contagious respiratory disease of chickens caused by chicken infectious bronchitis virus (IBV), and chickens of different ages, genders, and breeds are susceptible. IBV mainly infects the respiratory tract, intestinal tract, fallopian tubes and kidneys of chickens, which can cause lesions and damages to corresponding organs. More importantly, it is easy to cause secondary infections of bacteria and other pathogens, causing greater losses. It is o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63C12N15/66A61K48/00A61P31/14
Inventor 韦平李孟何坤
Owner GUANGXI UNIV
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