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Polypeptide for inhibiting porcine epidemic diarrhea virus (PEDV) infection, and application of polypeptide

A technology for porcine epidemic diarrhea and virus infection, which is applied in the fields of bioinformatics, cell biology and virology, and can solve problems such as anti-virus infection

Inactive Publication Date: 2019-03-12
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there have been antiviral studies on HR2 of other coronaviruses, antiviral infection based on PEDV HR2 has yet to be fully elucidated

Method used

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  • Polypeptide for inhibiting porcine epidemic diarrhea virus (PEDV) infection, and application of polypeptide
  • Polypeptide for inhibiting porcine epidemic diarrhea virus (PEDV) infection, and application of polypeptide
  • Polypeptide for inhibiting porcine epidemic diarrhea virus (PEDV) infection, and application of polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1. PEDV HR2 Prediction

[0022] HR1 and HR2 of S protein of PEDV CH / hubei / 2016 strain were predicted by online software LearnCoil-VMF (http: / / night-ingale.lcs.mit.edu / cgi-bin / vmf). The LearnCoil-VMF software recognizes typical HR features, that is, there are about 3 to 4 heptavalent repeats in HR1 and HR2 respectively, and each repeat starts from leucine or isoleucine. The result is as figure 1 As shown, the predicted HR1 and HR2 of PEDV S protein are located at amino acids 982-1121 and amino acids 1278-1317, respectively.

Embodiment 2

[0023] Embodiment 2.PEDV HR2 derived polypeptide design

[0024] The human coronavirus NL63 (HCoV-NL63) HR1 and HR2 amino acid sequences that have been analyzed with reference to the fusion core crystal structure ( figure 2 , wide frame), PEDV HR1 and HR2-derived polypeptides were designed, and finally amino acids 1005-1061 and 1273-1314 were selected as HR1 and HR2-derived polypeptide sequences, and the amino acid sequences were SAIGNITSAFESVKEAISQTSKGLNTVAHALTKVQEVVNSQGAALTQLTVQLQHNFQ (SEQ ID NO.1) and NATYLNLTGEIADLEQRSESLRNTTEELQSLIYNINNTLVDL (SEQ ID NO. 2). After analysis, its amino acid composition conforms to the characteristics of the seven-valent repeat of coronavirus, that is, a group of neatly arranged seven-valent repeat amino acid residues, and its residue positions can be represented by a, b, c, d, e, f and g, where a and The amino acid residue at the d position is a hydrophobic amino acid or an amino acid with a large side chain ( image 3 ). The HR1 and HR2...

Embodiment 3

[0025] Example 3. PEDV HR2-derived polypeptide inhibits PEDV from infecting host cells

[0026] 3.1 PEDV HR2-derived peptides inhibit the proliferation of PEDV

[0027] The PEDV host small intestinal epithelial cells IPEC-J2 were divided into 2 × 10 5 cells / mL, 500 μL / well for 24-well plate, at 37°C CO 2 Cultivate in the incubator for 24h. PEDV CH / hubei / 2016 strain and HR1, HR2 synthetically derived polypeptides were respectively diluted in serum-free DMEM medium (Gibco, USA). The multiplicity of infection (MOI) of the virus was 1. DMEM medium containing virus and dimethyl sulfoxide (DMSO) was used as control. After mixing the virus with MOI=1 and the synthetically derived polypeptide with a final concentration of 0.1-1000 nM, place it at 37° C. for 1 hour. The plate was washed 3 times with PBS, 500 μL of the mixed solution was added to each well, and three replicate wells were set up for each group of treated cells. After incubating at 37°C for 1 h, wash the plate three ...

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Abstract

The invention discloses a polypeptide for inhibiting porcine epidemic diarrhea virus (PEDV) infection, and application of the polypeptide. An amino acid sequence of the polypeptide is NATYLNLTGEIADLEQRSESLRNTTEELQSLIYNINNTLVDL. A PEDV heptad repeat 2 (PEDV HR2) is predicted based on bioinformatics, and the HR2-derived polypeptide is designed and synthesized by means of amino acid sequence alignment. The synthesized HR2-derived polypeptide is used for inhibiting an IPEC-J2 process of PEDV-infected host intestinal epithelial cells; after being subjected to a fluorescent quantitative polymerase chain reaction (PCR), the synthesized polypeptide has a function of inhibiting virus infection of host cells, and can be applied to resisting the PEDV infection. The polypeptide information provided bythe invention enriches the functions of a PEDV fusion core, and provides a reference for the subsequent study of a PEDV membrane fusion mechanism; the polypeptide provided by the invention can be applied to the preparation of antiviral vaccines and the medicines for PEDV.

Description

technical field [0001] The invention relates to the fields of virology, bioinformatics and cell biology, in particular to a polypeptide for inhibiting porcine epidemic diarrhea virus (porcine epidemic diarrhea virus, PEDV) infection and application thereof. Background technique [0002] Porcine epidemic diarrhea (PED) is a highly contagious porcine intestinal infectious disease, characterized by vomiting, watery diarrhea and dehydration in affected pigs. PED affects pigs of all ages, especially for suckling piglets within 7 days of age, with a mortality rate as high as 100%. PED has caused huge economic losses to my country's pig industry, and has become one of the infectious diseases that restrict the healthy development of domestic pig industry. PED pathogenic porcine epidemic diarrhea virus (PEDvirus, PEDV) is a single-stranded positive-sense RNA virus with an envelope, which belongs to the genus Alphacoronavirus (Alphacoronavirus) of the family Coronaviridae of the orde...

Claims

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Application Information

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IPC IPC(8): C07K14/165A61K39/215A61P31/14
CPCA61K39/12A61K2039/552A61P31/14C07K14/005C12N2770/20022
Inventor 张改平李睿孙彦刚乔松林陈鑫鑫郭振华卢清侠赵东李学伍
Owner HENAN ACAD OF AGRI SCI
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