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Multivalent recombinant protein subunit vaccine in chicken coccidiosis and application thereof

A technology of subunit vaccine and recombinant protein, applied in the field of chicken coccidia multivalent recombinant protein subunit vaccine

Active Publication Date: 2013-11-13
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether the corresponding prokaryotic expressed recombinant protein can also provide effective immune protection is unknown, and a large number of experiments are needed to explore and verify

Method used

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  • Multivalent recombinant protein subunit vaccine in chicken coccidiosis and application thereof
  • Multivalent recombinant protein subunit vaccine in chicken coccidiosis and application thereof
  • Multivalent recombinant protein subunit vaccine in chicken coccidiosis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1. Expression and purification of chicken coccidia TA4, NA4, LDH and Em8 recombinant proteins

[0026] 1. Construction of E.maximaEm8 gene prokaryotic expression plasmid

[0027] Respectively cut pMD18-T-Em8 with BamH I and Not I (Lei Chenyu. Immunogenicity study of Eimeria maxima antigen EmTFP250. Nanjing Agricultural University master's degree thesis, 2008) and pET32a(+) plasmid , the gel recovery kit was used to recover the target gene and pET32a(+) large fragment, and ligated to construct the pET32a(+)-Em8 recombinant plasmid.

[0028] 2. Preparation of genetically engineered bacteria

[0029]The E.tenella recombinant plasmid pET28-TA4 (Li Xiangrui. Immunomodulatory DNA vaccine for the prevention of chicken coccidiosis. People's Republic of China invention patent, patent number: ZL200510124048.4), E.necatrix recombinant plasmid pET28a-NA4 (Li Xiangrui et al. Immunomodulatory DNA vaccine to prevent Eimeria poisoning in chickens. Invention patent of the Peop...

Embodiment 2

[0032] Example 2. Detection of immune protection of chicken coccidia multivalent recombinant protein subunit vaccine

[0033] 1. Experimental Design

[0034] The 0-day-old chicks were raised in a strictly disinfected, coccidia-free environment and had free access to water and food. The 14-day-old experimental chickens were weighed one by one, and the overweight and underweight individuals were eliminated. They were randomly divided into groups and the average body weights between the experimental groups were adjusted to make them close to the same, with 30 chickens in each group. TA4, NA4, LDH and Em84 recombinant proteins were mixed in equal proportions, and chickens were immunized with the recombinant proteins to infect E. tenella, E. necatrix, E. acervulina and E. maxima respectively. A non-immune group infected with 4 kinds of coccidia (red control group) and a common non-infected non-immune group (white control group) were set up, a total of 9 groups.

[0035] In each e...

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Abstract

The invention relates to a technical filed of biological drugs and a multivalence recombinant protein subunit vaccine in chicken coccidiosis. The vaccine comprises four recombinant proteins in the chicken coccidiosis: TA4 (E.tenella), NA4 (E.necatrix), LDH (E.acervulina) and Em8 (E.maxima). It is proved through an animal immunity protective experiment that the vaccine can effectively resist four chicken coccidiosis infections and can be used in preparation of drug formulations for prevention against the chicken coccidiosis.

Description

technical field [0001] The invention relates to the technical field of biological veterinary medicine, and relates to a chicken coccidia multivalent recombinant protein subunit vaccine. Background technique [0002] Chicken coccidiosis is a parasitic protozoan disease that seriously endangers the production and development of the chicken industry, and the economic loss caused by it is as high as more than 2 billion pounds every year. With the increasingly serious problems of chicken coccidiostat resistance and drug residues, people expect to replace the use of anticoccidiostats with more ideal methods. However, the successful development and application of live coccidiosis vaccines provides a technical means that can replace drugs to control coccidiosis in chickens, but there are disadvantages such as high cost, difficulty in preservation, easy to disperse virus and threat of pathogenicity, among which the biggest disadvantage is It is easy to disperse the virus and has pat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/012A61P33/02
Inventor 李祥瑞严若峰徐立新宋小凯高云路
Owner NANJING AGRICULTURAL UNIVERSITY
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