DNA vaccine with resistance to infectious spleen and kidney necrosis viruses

A technology of spleen and kidney necrosis virus and DNA vaccine, which is applied in the field of bioengineering vaccines, and achieves the effects of simple preparation method, easy market promotion and application, and low cost

Inactive Publication Date: 2014-10-15
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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  • Claims
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  • DNA vaccine with resistance to infectious spleen and kidney necrosis viruses
  • DNA vaccine with resistance to infectious spleen and kidney necrosis viruses
  • DNA vaccine with resistance to infectious spleen and kidney necrosis viruses

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[0020] Experimental example 1:

[0021] 1. Construction of DNA vaccine

[0022] 1.1 Amplification of ISKNV MCP gene

[0023] (1) Design primers according to the MCP gene sequence of Infectious Spleen and Kidney Necrosis Virus (ISKNV) [HQ317460.1] in GenBank, and introduce restriction sites (marked underline). The primer sequences are as follows:

[0024] P1: 5’-CG GGTACC ATGGCTGCAATCTCA -3' (SEQ ID NO: 2), introduce KpnⅠ restriction site (underlined part),

[0025] P2: 5'-GC GAATTC TTACAGGATAGGGAAGC -3' (SEQ ID NO: 3), introducing EcoRI restriction site (underlined part), this pair of primers can amplify the MCP gene sequence (as shown in SEQ ID NO: 1).

[0026] (2) PCR amplification is carried out using the virus genome stored in our laboratory as a template. The 25μL PCR reaction system contains: 10×buffer (including Mg 2+ ) 2.5μL, 4×dNTP (10mmol / L) 0.5μL, upstream and downstream primers (20μmol / L) each 0.5μL, Taq DNA polymerase (5U / μL) 0.3μL, DNA 1.0μL, sterile double distilled wa...

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Abstract

The invention discloses a DNA vaccine with resistance to infectious spleen and kidney necrosis viruses (ISKNV). The nucleotide sequence of the DNA vaccine is the gene sequence of MCP (major capsid protein) of ISKNV. When the DNA vaccine is used together with an adjuvant QCDC, excellent immunogenicity is realized, the relative percent survival can reach 80% which is remarkably superior to the relative percent survival (64.3%) of the present MCP protein vaccine, and a novel vaccine with the resistance to ISKNV of siniperca chuatsi can be developed. A preparation method of the DNA vaccine is simple, easy to operate, low in cost and easy for popularization in the market.

Description

technical field [0001] The invention belongs to the field of bioengineering vaccines, and relates to a DNA vaccine, in particular to a DNA vaccine against infectious spleen and kidney necrosis virus (ISKNV). Background technique [0002] Siniperca chuatsi (Siniperca chuatsi) is one of the important freshwater fish cultured species in my country. The iridescent virus disease of mandarin fish caused by Infectious spleen and kidney necrosis virus (ISKNV) is the main disease of mandarin fish culture. The mortality rate of mandarin fish is close to 100%, which has become the main bottleneck restricting the development of mandarin fish aquaculture. For a long time, there has been no effective drug for the prevention and treatment of mandarin fish virus disease, and there are many disadvantages of drug prevention and control of diseases, such as pathogenic drug resistance, environmental pollution, food safety and so on. Therefore, as a disease control measure in line with the envir...

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Application Information

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IPC IPC(8): A61K48/00A61P31/20
Inventor 李宁求付小哲吴淑勤林强刘礼辉张德峰
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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