Chicken infectious laryngotracheitis bivalent nucleic acid vaccine and preparation method thereof

A nucleic acid vaccine, laryngotracheitis technology, applied in the directions of pharmaceutical formulations, genetic material components, introduction of foreign genetic material using carriers, etc. The effect of immune response strength, improving commission efficiency, and shortening incubation period

Inactive Publication Date: 2011-02-16
HENAN AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Attenuated vaccines have disadvantages such as strong virulence, latent infection, and easy reversion to ancestors. Long-term use of attenuated vaccines is not conducive to

Method used

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  • Chicken infectious laryngotracheitis bivalent nucleic acid vaccine and preparation method thereof
  • Chicken infectious laryngotracheitis bivalent nucleic acid vaccine and preparation method thereof
  • Chicken infectious laryngotracheitis bivalent nucleic acid vaccine and preparation method thereof

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[0048] Example

[0049] A bivalent nucleic acid vaccine for chicken infectious laryngotracheitis, in which the complete genes of ILTV HN1 strain gB and ChIL-18 are inserted into two multiple cloning sites of the eukaryotic expression vector pIRES. The bivalent nucleic acid vaccine co-expresses chicken Infectious laryngotracheitis virus gB gene and chicken IL-18 gene.

[0050] The preparation method of avian infectious laryngotracheitis bivalent nucleic acid vaccine, its steps are as follows:

[0051] (1) Design and synthesize a pair of primers to amplify ChIL-18,

[0052] The upstream primer Y1 is: 5’-GTA GGATCC ATGAGCTGTGA AGAGAT-3’,

[0053] Downstream primer Y2 is: 5’-TAC GTCGAC TTATAGGTTGTGCCTTTC-3’,

[0054] The upstream and downstream primers were added to the BamHI and Sal I restriction sites (underlined), and the expected amplification length is 615bp;

[0055] Design and synthesize a pair of primers to amplify ILTV gB,

[0056] The upstream primer Y3 is: 5’-GTTA CTCGAG ATGGCTAG...

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Abstract

The invention discloses a chicken infectious laryngotracheitis bivalent nucleic acid vaccine, wherein gB and ChIL-18 (Chicken Interleukin-18) complete genes of an ILTV (Infectious Laryngotracheitis Virus) HN1 strain are respectively inserted into two multiple cloning sites of an eukaryotic expression vector pIRES, and the chicken ILTV gB gene and the chicken IL-18 gene are co-expressed by the bivalent nucleic acid vaccine. The gB gene fragment and the ChIL-18 gene fragment of the chicken ILTV HN1 strain are respectively inserted into the multiple cloning enzyme-cutting sites of MCS A and MCS B at the downstream of a promoter of the eukaryotic expression vector pIRES adopted by the invention to obtain eukaryotic double-expression plasmids pIRES-gB/IL18. The invention avoids the problem of possible disproportionate cellular uptake when two plasmids are co-immunized, improves the local antigen presenting efficiency, enhances the local immune-response strength, shortens the immune-response latent period and lays a foundation for the research of novel ILTV vaccines.

Description

technical field [0001] The invention relates to a nucleic acid vaccine, in particular to a bivalent nucleic acid vaccine for chicken infectious laryngotracheitis and a preparation method thereof. Background technique [0002] Chicken infectious laryngotracheitis is an acute, highly contagious upper respiratory infectious disease. The disease is characterized by dyspnea, panting, coughing up bloody secretions, swelling and bleeding of the throat and tracheal mucosa, and the formation of erosions. During the acute outbreak of the disease, the morbidity rate can reach 100%, and the mortality rate is generally about 20%. Since the disease was reported in the United States in 1925, it has spread all over the world, and my country discovered the existence of the disease in the 1950s. [0003] At present, the main measure to prevent and control ILT at home and abroad is the use of attenuated vaccines. Attenuated vaccines have disadvantages such as strong virulence, latent infect...

Claims

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Application Information

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IPC IPC(8): A61K48/00C12N15/79A61P31/14
Inventor 陈红英崔保安魏战勇邵攀峰刘金朋王淑娟朱前磊王子馨钞安军
Owner HENAN AGRICULTURAL UNIVERSITY
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