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A kind of porcine circovirus type Ⅱ nucleic acid vaccine and its preparation method

A porcine circovirus and nucleic acid vaccine technology, which is applied in the directions of antiviral agents, viral antigen components, pharmaceutical formulations, etc., can solve the problems of low protection against viruses of DNA vaccines, shorten the incubation period of immune response, improve the efficiency of the extraction, and enhance the local The effect of immune response strength

Inactive Publication Date: 2011-12-14
HENAN AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0009] The technical problem to be solved in the present invention is that the DNA vaccine challenge protection of a single antigen of PCV2 is relatively low, and porcine interleukin-18 (pIL-18) is used as an important cellular immune adjuvant of the vaccine to prepare a pig interleukin-18 (pIL-18) Circovirus Type II Nucleic Acid Vaccine

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  • A kind of porcine circovirus type Ⅱ nucleic acid vaccine and its preparation method
  • A kind of porcine circovirus type Ⅱ nucleic acid vaccine and its preparation method
  • A kind of porcine circovirus type Ⅱ nucleic acid vaccine and its preparation method

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Embodiment Construction

[0042] A kind of porcine circovirus type II nucleic acid vaccine, porcine circovirus type II LY strain ORF2 and porcine IL-18 genes are respectively inserted into two multiple cloning sites of the eukaryotic expression vector pIRES, and the nucleic acid vaccine co-expresses porcine circovirus type II nucleic acid vaccine Circovirus type II LY strain Cap and porcine IL-18 proteins.

[0043] The preparation method of described porcine circovirus type II nucleic acid vaccine, its steps are as follows:

[0044] (1) Designed and synthesized primers pIL18fs / pIL18rs for porcine IL-18

[0045] The upstream primer pIL18fs is 5'-TAA GTC GAC ATGGCTGCTGAACCGGA-3',

[0046] The downstream primer pIL18rs is 5'-CGT GCGGCCGC TTAGTTTCTTGTTTTG-3',

[0047] The upstream and downstream primers were respectively added with Sal I and Not I restriction sites, and the amplified length was 599bp;

[0048] Designed and synthesized primers ORF2fs / ORF2rs for porcine ORF2

[0049] The upstream prim...

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Abstract

The technical scheme of the present invention is: a kind of porcine circovirus type II nucleic acid vaccine, respectively inserting porcine circovirus type II LY strain ORF2 and porcine IL-18 gene into two multiple cloning sites of the eukaryotic expression vector pIRES, The nucleic acid vaccine co-expresses porcine circovirus type II LY strain Cap and porcine IL-18 protein. The present invention is identified by PCR, double enzyme digestion and sequencing, which proves that the co-expression plasmid pIRES-ORF2 / PIL18 has been successfully constructed. The successfully constructed recombinant plasmid pIRES-ORF2 / PIL18 was transfected into porcine kidney PK-15 cells by liposome method, and the expression of the target protein was detected by RT-PCR and indirect immunofluorescence. The transcription of the two target genes was detected by RT-PCR; the specific bright green fluorescence of ORF2 protein was observed in the cytoplasm and nucleus by indirect immunofluorescence test, which proved that the protein was expressed.

Description

technical field [0001] The invention relates to a nucleic acid vaccine, in particular to a porcine circovirus type II nucleic acid vaccine and a preparation method thereof. Background technique [0002] Porcine circovirus (PCV) is the smallest animal virus discovered so far, and has two genotypes: porcine circovirus type I (PCV1) and porcine circovirus type II (PCV2). Pigs are susceptible to PCV2, and pigs of various ages and genders can be infected with PCV2. The common postweaning multisystemic wasting syndrome (Postweaning multisystemic wasting syndrome, PMWS) mainly occurs in piglets during lactation and rearing period, especially in piglets aged 5-12 weeks, and generally begins to develop symptoms 2-3 days after weaning. From the perspective of the season of onset, it can be onset all year round, but the peak period is in May, followed by April and June. The age of onset also varies with different regions, which is related to the unique predisposing factors, the level...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K39/12A61P31/20
Inventor 陈红英崔保安李新生魏战勇刘金朋王淑娟朱前磊王亚丹王子馨李坤
Owner HENAN AGRICULTURAL UNIVERSITY
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