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Specific binding peptide of duck hepatitis virus type I vp1 protein and its application
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A duck hepatitis virus, technology of binding peptides, applied in the direction of application, antiviral agents, peptides, etc.
Inactive Publication Date: 2016-08-31
HENAN UNIV OF SCI & TECH
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Problems solved by technology
The research on the new DHV-I is currently mostly focused on the analysis of the whole genome sequence, and there are few reports on the study of gene function
Method used
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Embodiment 1
[0023] The screening of duck hepatitis virus type I VP1 protein specific binding peptide in the present embodiment comprises the following steps:
[0024] (1) Preparation of DHV-1VP1 protein
[0025] 1) Construction of expression vector pET32a-VP1
[0026] According to the VP1 gene sequence (serial number: EF653378) of DHV-1 published by GenBank, use DNAStar software to analyze, design a pair of specific primers, primer F15' end introduces EcoR I restriction enzyme site, F25' end introduces Xba I restriction enzyme site. Primers were synthesized by Dalian Bao Biological Engineering Company.
[0027] The primer sequences are as follows:
[0028] F1:5'-CCAGAATTCGGTGATTCTAACCAG-3' (as shown in SEQ ID NO:2);
[0029] F2: 5'-GTTTCTAGATTCAATTTCCAG-3' (such as SEQ ID NO.3).
[0030] Duck hepatitis virus type I VJ strain is known to be isolated from a diseased duck farm, and determined to be type I duck hepatitis virus through sequence determination and evolution analysis, and it...
Embodiment 2
[0058] In this example, the application of duck hepatitis virus type I VP1 protein-specific binding peptide in the preparation of type I anti-duck hepatitis virus disease medicine includes the following steps: using solid phase synthesis to synthesize duck hepatitis virus type I VP1 protein specific binding peptide The peptide is purified, freeze-dried, and mixed with the commonly used auxiliary materials starch, sucrose, and talc in the medical field to obtain a tablet drug for resisting duck hepatitis virus disease. The duck hepatitis virus type I VP1 protein specific binding peptide is used as the main antiviral active ingredient in the antiviral drug, and the content of each tablet of the main drug is 0.1g, and the mass percentage of the main drug is 50%.
Embodiment 3
[0060] In this example, the application of duck hepatitis virus type I VP1 protein-specific binding peptide in the preparation of feed additives against duck hepatitis virus disease includes the following steps: purifying the duck hepatitis virus type I VP1 protein-specific binding peptide and freeze-drying, The polypeptide powder with a purity of over 95% is obtained. 0.2-0.5‰ duck hepatitis virus type I VP1 protein-specific binding peptide can be added according to the feed weight, that is, 200-500 grams of peptides can be added per ton of feed.
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Abstract
The invention discloses a duck hepatitis virus type I VP1 protein specific binding peptide and its application and belongs to the technical field of molecular biology of animal virus. According to the invention, DHV-1VP1 protein is obtained by a genetic engineering method; the DHV-1VP1 protein is used as a target; and bacteriophage random 12 peptide library screening is used to obtain DHV-1VP1 specific binding polypeptide with its amino acid sequence of LLSPGTHHSPWP. It proves through tests that the duck hepatitis virus type I VP1 protein specific binding peptide can inhibit proliferation of DHV-1 in duck embryo fibroblasts and different concentrations of the peptide can lower virus copy number of DHV-1 in the duck embryo fibroblasts remarkably and reduce virus titer of DHV-1 in the duck embryo fibroblasts substantially. The duck hepatitis virus type I VP1 protein specific binding peptide has a good application prospect in preparation of medicines or feed additives for resisting duck hepatitis virus and can be used for preventing and curing duck hepatitis virus.
Description
technical field [0001] The invention relates to a duck hepatitis virus type I VP1 protein specific binding peptide, and also relates to the application of the specific binding peptide, which belongs to the technical field of animal virus molecular biology. Background technique [0002] Duck viral hepatitis, also known as "back neck disease", is an acute and highly pathogenic infectious disease caused by duck hepatitis virus (DHV) that causes liver damage in ducklings. It is characterized by acute onset, rapid spread, and high mortality , The clinical manifestations of sick ducks are depression, neurological symptoms, movement disorders, opisthotonos, and the main pathological changes are hepatitis and hemorrhage. Duck viral hepatitis was first discovered in the United States, and the virus was isolated from chicken embryos, and later found in many duck-raising countries such as the United Kingdom, Canada, and Germany. Most provinces and regions in my country also have the o...
Claims
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Application Information
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