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Duck tembusu virus envelop E protein inhibitory peptide and application thereof

A duck Tembusu virus, protein inhibition technology, applied in the direction of application, antiviral agent, peptide/protein composition, etc.

Inactive Publication Date: 2014-12-10
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For this disease, there is no report abroad

Method used

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  • Duck tembusu virus envelop E protein inhibitory peptide and application thereof
  • Duck tembusu virus envelop E protein inhibitory peptide and application thereof
  • Duck tembusu virus envelop E protein inhibitory peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] In the present embodiment, the screening of duck Tembusu virus envelope E protein inhibitory peptide comprises the following steps:

[0023] 1) Preparation of DTMUV HN1 strain envelope E protein by conventional method

[0024] According to the gene sequence of the E protein of DTMUV HN1 strain published by GenBank (serial number: JQ669731), use DNAStar software to analyze, design a pair of specific primers P1 and P2, and add restriction enzyme sites EcoR I and Xba I to both ends of the primers respectively and protective bases (synthesized by Takara Company), the underlined part is the enzyme cleavage site.

[0025] Upstream primer P1: 5'-CCG GAATTC TTCAGCTGTCTGGGGATGCA-3' (such as SEQ ID NO: 2, the underlined part is the EcoR I restriction site);

[0026] Downstream primer P2: 5'-CGA TCTAGA ATTGACATTTACTGCCAGG-3' (such as SEQ ID NO: 3, the underlined part is the Xba I restriction site).

[0027] Conventional methods extract DTMUV HN1 strain RNA (viral RNA extra...

Embodiment 2

[0055] In this embodiment, the application of duck Tembusu virus envelope E protein inhibitory peptide in the preparation of medicines against duck Tembusu virus disease includes the following steps: take the duck Tembusu virus envelope E protein inhibitory peptide after purification and freeze-drying powder (duck Tembusu virus envelope E protein inhibitory peptide is synthesized by solid-phase peptide synthesis, purified by reverse high-performance liquid chromatography, and the purity after purification and freeze-drying reaches more than 95%), and commonly used starch, sucrose, Talcum powder is mixed to obtain the tablet medicine for inhibiting the proliferation of duck Tembusu virus. The duck Tembusu virus envelope E 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 0.1g. The percentage content is 50%. Alternatively, the peptide powder is dissolved in PBS buffer to prepare peptid...

Embodiment 3

[0057] In this embodiment, the application of the duck Tembusu virus envelope E protein inhibitory peptide in the preparation of feed additives for inhibiting the duck Tembusu virus proliferation comprises the following steps: freezing the duck Tembusu virus envelope E protein inhibitory peptide after purification dry to obtain a polypeptide powder with a purity of more than 95% (purification adopts high performance liquid chromatography, and freeze-drying adopts freeze-drying method). 0.2-0.5‰ duck Tembusu virus envelope E protein inhibitory peptide can be added according to the feed weight, that is, 200-500 grams of peptide powder can be added per ton of feed.

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Abstract

The invention discloses a duck tembusu virus envelop E protein inhibitory peptide and application thereof, belonging to the technical field of animal virus molecular biologics. DTMUV envelop E protein is obtained by using a genetic engineering method, DTMUV HN1 strain E protein is taken as a target, the duck tembusu virus envelop E protein inhibitory peptide is obtained through screening by using a bacteriophage random 12 peptide library, and the amino acid sequence of the duck tembusu virus envelop E protein inhibitory peptide is as shown in HWSTRQGSTRWN. By adopting the inhibitory peptide, proliferation of the duck tembusu virus in duck embryo fibroblast can be inhibited, the virus copy number of the DTMUV in the duck embryo fibroblast can be remarkably reduced in different concentrations, meanwhile the virus titer of the DTMUV in the duck embryo fibroblast can be remarkably reduced, and the DTMUV has no remarkable influence on proliferation of the duck embryo fibroblast. The duck tembusu virus envelop E protein inhibitory peptide has good application prospect in preparing medicines or feed additives for preventing the duck tembusu virus, and can be used for preventing and controlling the duck tembusu virus.

Description

technical field [0001] The invention relates to a duck Tembusu virus envelope E 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] In recent years, a new viral infectious disease of ducks has broken out in southern China, mainly causing a serious decline in duck egg production. The disease spreads rapidly and involves a wide range. For paralysis, rolling, unsteady standing and ataxia, some sick ducks even died within a few days. Autopsy dead ducks, most of them showed obvious enlarged spleen and kidney, and laying ducks showed hemorrhagic oophoritis. The disease first broke out in Jiangsu and Zhejiang, then spread to Fujian and Anhui, and then successively broke out in Jiangxi, Shandong, Hunan, Henan, and Hebei. For this disease, foreign countries have not yet been reported. Domestic researchers have isolated the pathogen, observed ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K38/10A61P31/14A61P35/00A23K1/16
Inventor 王臣李小康郭香玲汪洋牛明媚吴庭才
Owner HENAN UNIV OF SCI & TECH
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