Preparation and Application of Bivalent DNA Vaccine for Prevention of White Spot Syndrome Virus in Shrimp

A technology of white spot syndrome and DNA vaccine, which is applied in the fields of application, antiviral agent, and medical preparations containing active ingredients, and can solve the problems of virus preservation and multiplication difficulties, etc.

Active Publication Date: 2019-01-01
湛江出入境检验检疫局检验检疫技术中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no cell line that can be used for the propagation of shrimp white spot syndrome virus, so it is very difficult to preserve and propagate the virus, so it is also facing great challenges in the development of vaccines

Method used

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  • Preparation and Application of Bivalent DNA Vaccine for Prevention of White Spot Syndrome Virus in Shrimp
  • Preparation and Application of Bivalent DNA Vaccine for Prevention of White Spot Syndrome Virus in Shrimp
  • Preparation and Application of Bivalent DNA Vaccine for Prevention of White Spot Syndrome Virus in Shrimp

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] The preparation method of preventing prawn white spot syndrome virus bivalent DNA vaccine, the steps are as follows:

[0074] Step 1: Design primers for the amplification of white spot virus VP19 and VP28 genes, as follows:

[0075] 1. Based on the VP19 gene sequence, use Primer Premier 5 software to synthesize a pair of primers:

[0076] (1-1) Upstream primer: 5- GGATCC ATGGCCACCACGACTAACAC-3, where the underlined GGATCC for BamHI;

[0077] (1-2) Downstream primer: 5- GAATTC TTACTGCCTCCTCTTGGGGT-3, where the underlined GAATTC is EcoRI;

[0078] 2. According to the VP28 gene sequence, use Primer Premier 5 software to synthesize a pair of primers:

[0079] (2-1) Upstream primer: 5-AGA GAATTC ATGGATCTTTCTTTCAC-3, where the underlined GAATTC is EcoRI;

[0080] (2-2) Downstream primer: 5-CAC AAGCTT TTACTCGGTCTCAGTGC-3, where the underlined AAGCTT is XhoI;

[0081] Step 2: Extraction of total white spot virus DNA, as follows:

[0082] Use the DNA extractio...

Embodiment 2

[0124] The preparation method of preventing prawn white spot syndrome virus bivalent DNA vaccine, the steps are as follows:

[0125] Step 1: Design primers for the amplification of white spot virus VP19 and VP28 genes, as follows:

[0126] 1. Based on the VP19 gene sequence, use Primer Premier 5 software to synthesize a pair of primers:

[0127] (1-1) Upstream primer: 5- GGATCC ATGGCCACCACGACTAACAC-3, where the underlined GGATCC for BamHI;

[0128] (1-2) Downstream primer: 5- GAATTC TTACTGCCTCCTCTTGGGGT-3, where the underlined GAATTC is EcoRI;

[0129] 2. According to the VP28 gene sequence, use Primer Premier 5 software to synthesize a pair of primers:

[0130] (2-1) Upstream primer: 5-AGA GAATTC ATGGATCTTTCTTTCAC-3, where the underlined GAATTC is EcoRI;

[0131] (2-2) Downstream primer: 5-CAC AAGCTT TTACTCGGTCTCAGTGC-3, where the underlined AAGCTT is XhoI;

[0132] Step 2: Extraction of total white spot virus DNA, as follows:

[0133] Use the DNA extractio...

Embodiment 3

[0175] The preparation method of preventing prawn white spot syndrome virus bivalent DNA vaccine, the steps are as follows:

[0176] Step 1: Design primers for the amplification of white spot virus VP19 and VP28 genes, as follows:

[0177] 1. Based on the VP19 gene sequence, use Primer Premier 5 software to synthesize a pair of primers:

[0178] (1-1) Upstream primer: 5- GGATCC ATGGCCACCACGACTAACAC-3, where the underlined GGATCC for BamHI;

[0179] (1-2) Downstream primer: 5- GAATTC TTACTGCCTCCTCTTGGGGT-3, where the underlined GAATTC is EcoRI;

[0180] 2. According to the VP28 gene sequence, use Primer Premier 5 software to synthesize a pair of primers:

[0181] (2-1) Upstream primer: 5-AGA GAATTC ATGGATCTTTCTTTCAC-3, where the underlined GAATTC is EcoRI;

[0182] (2-2) Downstream primer: 5-CAC AAGCTT TTACTCGGTCTCAGTGC-3, where the underlined AAGCTT is XhoI;

[0183] Step 2: Extraction of total white spot virus DNA, as follows:

[0184] Use the DNA extractio...

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Abstract

The preparation method and application of the bivalent DNA vaccine for preventing prawn white spot syndrome virus of the present invention are carried out in the design of primers. Double enzyme cutting site; when performing double enzyme cutting, the target gene can obtain sticky ends, and then use DNA ligase to serially clone the antigenic genes VP19 and VP28 into the prokaryotic expression vector for prokaryotic expression vector; in the construction of bivalent DNA For vaccines, VP19 gene modification is required, that is, the stop codon of the VP19 gene is removed through primer design; since the restriction site downstream of the VP19 gene is the same as the restriction site upstream of VP28 during primer design, the The VP19 gene and the VP28 gene can be cloned in tandem by enzyme cutting technology.

Description

technical field [0001] The invention belongs to the technical field of DNA vaccines, in particular to a preparation method and application of a bivalent DNA vaccine for preventing prawn white spot syndrome virus, and in particular to the technical field of biological veterinary medicine. The vaccine contains white spot syndrome virus (white spot syndrome virus) , WSSV) envelope protein VP19 antigen gene and envelope protein VP28 antigen gene. At the same time, the vaccine contains PVAX1.0 vector. Background technique [0002] Shrimp white spot syndrome (White spot syndrome) is a fulminant infectious disease of prawns caused by white spot syndrome virus (White spot syndrome Virus, WSSV). Farming areas have caused huge economic losses. WSSV is a kind of enveloped non-inclusion body baculovirus, which belongs to the Nimaviridea family. It is a circular double-stranded DNA virus with an envelope. The DNA of WSSV virus is 290KD, with 181 ORFs, including 39 structural proteins. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K39/295A23K20/00A23K50/80A61P31/20
Inventor 张娜马新华龙阳谢艳辉袁俊杰刘骁杨劲李红权徐金龙孙良娟
Owner 湛江出入境检验检疫局检验检疫技术中心
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