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Bird infectious bronchitis virus N-gene and bird IL-2 gene co-expressing DNA bacterin pIRES-N/IL2

A technology of DNA vaccine and bronchitis, which is applied in the field of animal medicine and bioengineering, can solve the problems of high cost, complicated preparation, and large dosage

Inactive Publication Date: 2008-09-03
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage is that the dosage is large, and adjuvants are needed, and the preparation is more complicated, so the cost is higher
However, if the DNA vaccine constructed separately and the GM-CSF encoding plasmid are mixed and injected, it will interfere with genetic immunity (Kamath AT, Hanke T, et al. Immuology, 1999, 96: 511; Hong Xiaowu, Dou Jun, etc., practical Journal of Oncology, 2003, 18: 154)

Method used

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Embodiment Construction

[0008] 1. The eukaryotic expression plasmids pIBVN, pDNAIL-2 and eukaryotic expression vector pIRES-EGFP / DsRed used in the present invention are constructed and provided by our laboratory; the host Escherichia coli JM109 is preserved by our laboratory; avian infectious bronchitis virus challenge The strain SAIBk used was identified and preserved by the laboratory.

[0009] 2. The design and synthesis of primers were designed after multiple comparisons based on chicken interleukin-2 gene sequences registered in GenBank at home and abroad. According to the restriction site on the eukaryotic expression vector pIRES-EGFP / DsRed, the upstream primer is designed at the start codon, and the BamHI restriction site is added; the downstream primer is designed at the end of the gene, and the NotI restriction site is added point.

[0010] The primer sequences were: upstream primer: 5'-CCAGGATCCACCATGATGTGCAAAG-3'; downstream primer: 5'-GAAGCGGCCGCAGATTAGTTAGC-3'.

[0011] 3. Construction...

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Abstract

The invention pertains to the research field of animal pharmaceutical and biological engineering, which is a DNA vaccine pIRES-N / IL2 co-expressed by N gene of an bird infectious bronchitis virus and bird IL-2 gene. pIBVN, pIRES-EGFP / DsRed plasmid is double digested with NheI and XhoI, N gene and carrier segment are gel recovered and connected together, and eucaryon expression plasmid pIRES-N / DsRed is constructed. Bird original IL-2 gene is PCR obtained using pDNAIL-2 plasmid as template. After that plasmid of IL-2, pIRES-N / DsRed are double digested by BamHI and NotI, IL-2 gene and carrier segment is gel recovered and connected, and DNA vaccine pIRES-N / IL2 co-expressed by N gene of Chinese isolate SAIBk of bird infectious bronchitis virus and bird IL-2 gene is constructed. The vaccine can be used in preventing bird infectious bronchitis.

Description

[technical field] [0001] The invention relates to the field of animal medicine bioengineering, and relates to a DNA vaccine PIRES-N / IL2 co-expressed with avian infectious bronchitis virus N gene and avian IL-2 gene. [Background technique] [0002] Avian Infectious Bronchitis (IB) is an acute, highly contagious infectious disease of chickens caused by the Avian Infectious Bronchitis virus of the Coronaviridae family Coronaviridae, characterized by respiratory symptoms, Decreased egg production quantity and quality, kidney disease, etc. Avian infectious bronchitis virus can infect chickens of all ages, leading to growth retardation, death, weight gain and reduced feed remuneration; it also often causes secondary infections such as mycoplasma infection and colibacillosis to increase the mortality of chicken flocks, Bring huge loss to chicken production. At present, the disease is widely prevalent in the world and is one of the major infectious diseases that seriously endanger...

Claims

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Application Information

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IPC IPC(8): C12N15/79C12N15/62A61K48/00A61P31/14A61P11/00
Inventor 王红宁阳泰张毅李玉玲唐梦君夏青青曾瑜虹
Owner SICHUAN UNIV
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