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Research method for restraining porcine epidemic diarrhea virus infection based on integrin ligand recognition peptides

A technology for porcine epidemic diarrhea and virus infection, applied in the field of porcine epidemic diarrhea virus infection, to achieve the effects of preventing virus infection, simple processing technology, and low production cost

Active Publication Date: 2020-08-25
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, small molecular compounds designed to target integrins may inhibit PEDV infection

Method used

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  • Research method for restraining porcine epidemic diarrhea virus infection based on integrin ligand recognition peptides
  • Research method for restraining porcine epidemic diarrhea virus infection based on integrin ligand recognition peptides
  • Research method for restraining porcine epidemic diarrhea virus infection based on integrin ligand recognition peptides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] A research method for inhibiting porcine epidemic diarrhea virus infection based on an integrin ligand recognition peptide, comprising the following steps,

[0047] S1: Dilute Vero E6 cell suspension grown and maintained in high-glucose DMEM medium containing 10% fetal bovine serum and 1% penicillin-streptomycin solution to 1×10 5 pc / mL, inserted into 100cm 2 Cell culture dish, shake gently to distribute cells evenly, then place in 5% CO 2 , cultured in a 37°C cell culture incubator;

[0048] S2: when the density of Vero E6 cells reaches 90%, the culture medium is discarded, and the Vero E6 cells are rinsed;

[0049] Specifically, Vero E6 cells were gently rinsed three times with PBS, and then rinsed once with serum-free DMEM basal medium.

[0050] S3: Add the rinsed Vero E6 cells into the basal medium containing 40 μmol integrin ligand recognition peptide, and incubate in a cell culture incubator at 37°C for 6 hours;

[0051] Specifically, the integrin ligand recog...

Embodiment 2

[0077] The difference between Example 2 and Example 1 is that the integrin ligand recognition peptide used is com, and other operations are the same.

Embodiment 3

[0079] The difference between Example 3 and Example 1 is that the integrin ligand recognition peptide used is er, and other operations are the same.

[0080] Further, the research results in Example 1 and Example 3 are as follows:

[0081] 1. Analysis of integrin ligand recognition sequence in PEDV S protein

[0082] Import the PEDV CV777 strain S protein amino acid sequence (GenBank: AF353511.1) downloaded from the NCBI database into the Editseq software to search for the integrin ligand recognition sequence, as shown in Table 3. The integrin ligand recognition sequence DGE is located in Amino acids 1133-1135, KGE at amino acids 621-623 and 655-657, RLD at amino acids 1070-1072, IET at amino acids 67-69, LDV at amino acids 643-645 and 1266 -1268 amino acids, IDS is located at 81-83 amino acids.

[0083] Table 3 The position of integrin ligand recognition sequence in PEDV S protein

[0084]

[0085] Download the S protein model (PDB: 6U7K) of PEDV classic strain CV777 in...

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Abstract

The invention discloses a research method for restraining porcine epidemic diarrhea virus infection based on integrin ligand recognition peptides. The method comprises the following steps of S1, diluting a Vero E6 cell suspension, and placing the diluted Vero E6 cell suspension in a cell culture incubator for culturing; S2, when cell density achieves 90%, removing a culture medium, and performingmoistening; S3, adding the cells to a basic culture medium containing the integrin ligand recognition peptides, and performing hatching; S4, after the liquid is removed, adding virus maintaining liquid; S5, performing inoculating with porcine epidemic diarrhea viruses; S6, after the Vero E6 cells inoculated with the viruses are placed in the cell culture incubator for 2h, sucking liquid; S7, adding the virus maintaining liquid, and placing mixed liquid in the cell culture incubator for culturing; S8, storing cells; S9, collecting samples, and performing real-time fluorescent quantitation PCR;S10, collecting samples, and performing western blotting; and S11, counting research results. The research method can verify whether the integrin ligand recognition peptides est, com and er have the effect of resisting PEDV infection, and reference is provided for prevention and control of PED and development of antiviral drugs.

Description

technical field [0001] The invention relates to the technical field of porcine epidemic diarrhea virus infection, in particular to a research method for inhibiting porcine epidemic diarrhea virus infection based on an integrin ligand recognition peptide. Background technique [0002] Porcine epidemic diarrhea (porcine epidemic diarrhea, PED) is caused by porcine epidemic diarrhea virus (porcine epidemic diarrhea virus, PEDV), a highly contagious porcine intestinal infectious disease characterized by diarrhea, vomiting, and dehydration. The lethality rate of suckling piglets is as high as 100%. PEDV belongs to Nidoviridae, Coronaviridae, Alphacoronavirus (Alphacoronavirus) members, is an enveloped, non-segmented, single-stranded positive-sense RNA virus. In recent years, PEDV infection has shown an obvious upward trend. In 1972, PED was first reported in the UK. After 2010, large-scale outbreaks occurred in China, South Korea, the Philippines, the United States, Germany an...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851G01N33/577G01N33/569
CPCC12Q1/701C12Q1/6851G01N33/577G01N33/56983G01N2333/183C12Q2561/113C12Q2563/107C12Q2531/113Y02A50/30
Inventor 孙东波李春秋张金悦苏明俊齐珊珊
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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