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.
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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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