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Grouper iridovirus recombinant envelope protein vaccine and application thereof

An iridescent virus and envelope protein technology, which is applied in the fields of molecular biology and immunology to achieve the effect of high protection rate, improved immune protection effect and low cost

Inactive Publication Date: 2018-07-13
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Envelope protein is an important surface antigen of enveloped iridescent virus, and there is currently no report of recombinant envelope protein vaccine against Singaporean grouper iridescent virus SGIV

Method used

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  • Grouper iridovirus recombinant envelope protein vaccine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The immunoprotective recombinant protein vaccine (recombinant envelope protein vaccine of grouper iridescent virus) is encoded by the nucleotide sequence shown in SEQID NO.1 in the sequence table.

[0032] The preparation method of immunoprotective recombinant protein vaccine (recombinant envelope protein vaccine of grouper iridescent virus):

[0033] 1) Construction of plasmid pET32a-VP88

[0034]Plasmid pET32 (Novagen, USA) was digested with EcoR I / Sal I, and a 5.9 kb fragment was recovered. Using the independently isolated and preserved Singapore grouper iridescent virus SGIV DNA as a template, use primer VP88-F: 5'-GCGAATTCATGGGCGCAGCGCAATC-3', primer VP88-R: 5'-GCGTCGACTCACTTTGCAGCTTCTC-3' to perform PCR under the following conditions: 95°C The template DNA was pre-denatured for 180s, followed by 35 cycles at 94°C for 30s, 55°C for 90s, and 72°C for 60s, and then extended at 72°C for 10 minutes. After the PCR product was purified, it was digested with EcoR I / Sal ...

Embodiment 2

[0038] Immune Application of Recombinant Protein Vaccine

[0039] Step 1): Preparation of recombinant protein vaccine and adjuvant mixture.

[0040] Recombinant protein vaccine mixture: the recombinant protein vaccine rVP88 (200mg / L, purity more than 90%) purified in Example 1 is adjusted to contain 200mg of bases in the sequence table SEQ ID NO.1 in every liter of buffer solution with PBS buffer Sequence coded vaccine protein solution (i.e. containing 200mg recombinant protein vaccine rVP88 / L) to obtain recombinant protein vaccine mixture;

[0041] Recombinant protein vaccine and adjuvant mixture: mix the recombinant protein vaccine rVP88 (200mg / L, with a purity of more than 90%) purified in Example 1 with aluminum hydroxide solution, and adjust it to contain 200mg per liter of buffer with PBS buffer The vaccine protein (recombinant protein rVP88) encoded by the base sequence in the sequence listing SEQ ID NO.1 and 0.1mg / 8 mL of a mixed solution of aluminum hydroxide (that...

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Abstract

The invention discloses a grouper iridovirus vaccine and a preparation method and an application of the grouper iridovirus vaccine. The grouper iridovirus vaccine is a protein encoded by the nucleotide sequence shown as in SEQ ID NO.1. The grouper iridovirus vaccine is prepared by constructing a plasmid pET32a-VP88, transforming escherichia coli BL21 (DE3), inducing the expression, breaking a cell, and recovering a recombinant protein from a supernatant. The recombinant protein rVP88 is mixed with an adjuvant, and a vaccine mixed solution has an immunoprotection effect on the grouper iridovirus SGIV. The grouper iridovirus vaccine has a stable immune effect, is higher in protection rate, is efficient in protection, and can be applied to protecting the fish fries and adult fish against theinfection of the grouper iridovirus in the cultivation process of groupers.

Description

Technical field: [0001] The invention belongs to the fields of molecular biology and immunology, and in particular relates to an immunoprotective Singapore grouper iridescent virus recombinant envelope protein vaccine and a preparation method and application thereof. Background technique: [0002] Grouper is a marine fish with high economic value. In recent years, the scale of coastal grouper farming has been expanded, but the breeding environment is deteriorating day by day, and various diseases, especially viral diseases, frequently break out, causing significant economic losses. Iridescent virus is an important pathogenic agent of grouper, which can cause fish mortality from 30% (in adult fish) to 100% (juvenile stage). In Asia, especially when the water temperature is above 25°C, this virus is very likely to infect rare seawater fish cultured in East Asia and South Asia, such as grouper, barramundi, red sea bream, flounder and large yellow croaker, etc. , resulting in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61P31/20C12N15/34C12N15/70C07K14/01
CPCA61K39/12A61K2039/552A61K2039/55505C07K14/005C12N15/70C12N2710/00022C12N2710/00034
Inventor 秦启伟周胜黄晓红黄友华魏京广
Owner SOUTH CHINA AGRI UNIV
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