Newcastle disease virus and duck circovirus fusion protein as well as coding gene and application thereof

A technology of duck circovirus and Newcastle disease virus, applied in gene therapy, antiviral agents, virus antigen components, etc., can solve the problems of DNA vaccines that have not been reported

Active Publication Date: 2014-01-22
GUANGXI VETERINARY RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, DNA vaccines targeting the fusion expression of the two viruses have not been reported at home and abroad.

Method used

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  • Newcastle disease virus and duck circovirus fusion protein as well as coding gene and application thereof
  • Newcastle disease virus and duck circovirus fusion protein as well as coding gene and application thereof
  • Newcastle disease virus and duck circovirus fusion protein as well as coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1. Construction and expression identification of recombinant eukaryotic expression vector pcDNA3.1-F-Cap

[0042]1. Acquisition of the F gene of duck-derived Newcastle disease virus and the Cap gene of duck circovirus

[0043] 1. Design and synthesis of primers

[0044] According to the F gene sequence of duck Newcastle disease virus (DuNDV) and the Cap gene sequence of duck circovirus (DuCV), primers were designed and synthesized (Table 1). Among them, primers F1 and F2 were used to amplify the ORF of the F gene of duck-derived NDV, primers C1 and C2 were used to amplify the ORF of the Cap gene of DuCV, and the four primers carried HindⅢ, BamHI, BamHI and EcoRI restriction sites respectively ( part in italics). The linker is underlined, and the wavy line is the kozak sequence. Primers were synthesized by Shanghai Invitrogen Company.

[0045] The primer sequence of table 1F gene and Cap gene

[0046]

[0047] 2. Nucleic acid extraction and target gene am...

Embodiment 2

[0080] Embodiment 2, animal immunization and antibody detection

[0081] 1. Grouping and immunization of test ducks

[0082]40 two-week-old SPF ducklings were randomly divided into 4 groups, 10 in each group. Blood was collected from ducks in each group before inoculation, and serum was separated. In the first group, each SPF duck was immunized with 100 μg of the recombinant plasmid pcDNA3.1-F-Cap constructed in Example 1; in the second group, each SPF duck was immunized with 200 μg of the recombinant plasmid pcDNA3.1-F-Cap; in the third group, each SPF duck was immunized with recombination Plasmid pcDNA3.1-F-Cap300μg; the fourth group (empty vector control group) immunized each SPF duck with 300μg of pcDNA3.1(+) empty vector. Immunization was performed by multiple injections into the chest muscles. Two weeks after the first immunization, a booster immunization with the same dose as the first immunization was carried out, blood was collected two weeks after each immunizatio...

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Abstract

The invention discloses newcastle disease virus (NDV) and duck circovirus (DuCV) fusion protein as well as a coding gene and application thereof. The fusion protein provided by the invention sequentially contains F protein of duck source newcastle disease virus and Cap protein of duck circovirus from an amino terminal to a carboxyl terminal; the amino acid sequence of the F protein of the duck source newcastle disease virus is the 1st to 553rd bits of a sequence 1 in a sequence table; the amino acid sequence of the Cap protein of the duck circovirus is the 574th to 829th bits of the sequence 1 in the sequence table. According to the invention, a Cap gene of the duck circovirus and an F gene of a duck source NDV isolate are used for constructing a fusion expression recombinant eukaryotic plasmid pcDNA3.1-F-Cap for the first time, the fusion expression recombinant eukaryotic plasmid pcDNA3.1-F-Cap is used as a DNA vaccine to immune SPF ducklings of 2 weeks old, and the result shows that the DNA vaccine can effectively promote the generation of an NDV antibody and a DuCV antibody by the SPF ducklings. The invention provides a theoretical and practical basis for researching a gene vaccine of DuCV and duck source NDV.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a fusion protein of Newcastle disease virus and duck circovirus and its encoding gene and application thereof, in particular to a fusion protein formed by duck-derived Newcastle disease virus F protein and duck circovirus Cap protein and its coding genes and applications. Background technique [0002] Newcastle disease of duck origin is an acute, highly contagious and fatal infectious disease of ducks caused by Newcastle disease virus (NDV). Early studies at home and abroad have shown that poultry NDV never infects waterfowl, and virulent infection does not show obvious clinical symptoms. However, in recent years, it has been discovered that the natural prevalence of NDV from ducks, which causes high pathogenicity and mortality, not only causes great losses to the duck industry, but also poses a threat to the chicken industry in my country. The F protein of Newcastle disease virus is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/63C12N5/10C12N1/21C12N1/15C12N1/19A61K39/295A61K39/12A61K39/17A61K48/00A61K39/395A61P31/14A61P31/20
Inventor 谢芝勋许宗丽谢丽基刘加波邓显文谢志勤庞耀珊范晴罗思思
Owner GUANGXI VETERINARY RES INST
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