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Duck hepatitis virus type I vp1 protein inhibitory peptide and its application

A technology for duck hepatitis virus and protein inhibition, which is used in applications, antiviral agents, peptides, etc.

Inactive Publication Date: 2016-08-17
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in recent years, many scholars at home and abroad have isolated new DHV-1 strains from diseased duck populations. There are significant differences in sequence between the new DHV-1 and the previously isolated DHV-1, and the new DHV-1 has no Cross-reacts with DHV-1

Method used

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  • Duck hepatitis virus type I vp1 protein inhibitory peptide and its application
  • Duck hepatitis virus type I vp1 protein inhibitory peptide and its application
  • Duck hepatitis virus type I vp1 protein inhibitory peptide and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0023] The screening of duck hepatitis virus type I VP1 protein inhibitory 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 its geno...

Embodiment 2

[0058] In this embodiment, the application of duck hepatitis virus type I VP1 protein inhibitory peptide in the preparation of anti-duck hepatitis virus disease medicine includes the following steps: adopt solid phase synthesis to synthesize duck hepatitis virus type I VP1 protein inhibitory peptide, purify, freeze-dry , mixed with commonly used auxiliary materials starch, sucrose and talcum powder in the field of medicine to prepare tablet medicine for resisting duck hepatitis virus disease. The duck hepatitis virus type I VP1 protein inhibitory peptide is used as the main antiviral active ingredient in the antiviral drug, the content of each tablet of the main drug is 0.1 g, and the mass percentage of the main drug is 50%.

Embodiment 3

[0060] In this embodiment, the application of the duck hepatitis virus type I VP1 protein inhibitory peptide in the preparation of feed additives against duck hepatitis virus disease comprises the following steps: after the duck hepatitis virus type I VP1 protein inhibitory peptide is purified, it is freeze-dried to obtain a purity of 95 More than % of the polypeptide powder, to get. 0.2-0.5‰ duck hepatitis virus type I VP1 protein inhibitory 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 inhibitory peptide and application thereof, belonging to the technical field of animal virus molecular biology. The present invention obtains DHV-1VP1 protein through genetic engineering method, uses DHV-1VP1 protein as a target, and obtains an inhibitory peptide specifically binding to DHV-1VP1 by using phage random 12 peptide library screening, and its amino acid sequence is: LLADTTHHRPWT. Experiments have shown that duck hepatitis virus type I VP1 protein inhibitory peptide can inhibit the proliferation of DHV-1 in duck embryo fibroblasts, and can significantly down-regulate the viral copy number of DHV-1 in duck embryo fibroblasts at different concentrations, and at the same time The method can significantly reduce the virus titer of DHV‑1 in duck embryo fibroblasts, can be used to prepare anti-duck hepatitis virus disease medicine or feed additive, and has a good application prospect in the prevention and treatment of DHV‑1.

Description

technical field [0001] The invention relates to a duck hepatitis virus type I VP1 protein inhibitory peptide, and also relates to the application of the inhibitory 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 occurrence of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08A61K38/10A61P31/14A61P35/00A23K20/147
Inventor 王臣何雷李小康牛明媚张春杰
Owner HENAN UNIV OF SCI & TECH
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