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Protein bonded with rabbit hemorrhagic disease virus VP60 protein and application thereof

A rabbit hemorrhagic disease, combined technology, applied in the field of biomedicine

Inactive Publication Date: 2013-04-17
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Membrane protein (Moesin, MSN) plays an important role in the reproduction and transmission of parvovirus, retrovirus and lentivirus (AIDS virus), but whether MSN can participate in the reproduction and transmission of RHDV has not been reported yet

Method used

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  • Protein bonded with rabbit hemorrhagic disease virus VP60 protein and application thereof
  • Protein bonded with rabbit hemorrhagic disease virus VP60 protein and application thereof
  • Protein bonded with rabbit hemorrhagic disease virus VP60 protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 In vitro combination of moesin MSN protein and virion

[0043] RHDV virus particles have the characteristics of agglutinating human blood "O" type red blood cells. 1.15 μg of recombinant MSN protein and virus particles with 8 hemagglutination units, incubated at room temperature for 45 minutes, can inhibit the hemagglutination activity of the virus particles, while the control group BSA has no Hemagglutination inhibitory activity, the results are shown in Table 1, figure 1 shown.

[0044] Table 1 The hemagglutination inhibitory titer of recombinant membrane moesin MSN protein to RHDV (unit: μ g)

[0045]

Embodiment 2

[0046] Example 2 Moesin MSN protein virus neutralization test

[0047] Take 10 RHDV antibody-negative rabbits and divide them into two groups, 5 rabbits in each group. The first neutralization group: RHDV virions with 16 times the median lethal dose (LD50) were mixed with 56 μg of recombinant MSN protein, the mixture was incubated at room temperature for 60 minutes, and inoculated by oral or nasal drops. The second group of negative control group: direct inoculation of 16 times half lethal dose (LD50) of RHDV virus particles through oral cavity and nasal drop. During the observation period of 8 days, all the 5 rabbits in the neutralization group survived and were healthy in appearance, and there was no abnormality in each organ (such as figure 2 Shown), the human "O" red blood cell agglutination titer of the liver tissue grinding liquid is 20, which is judged as negative. 5 rabbits in the negative control group died one after another within 3 days, and the human "O" hemoglo...

experiment example 1

[0050] Experimental example 1 The protein that interacts with the structural protein VP60 of rabbit hemorrhagic disease virus was obtained from the cDNA library of rabbit liver tissue

[0051] The yeast two-hybrid method was used to catch the protein interacting with the VP60 protein of rabbit hemorrhagic disease virus. The oligonucleotide primers used in this study are shown in Table 3:

[0052] Primer name

Primer sequence

VP60 upstream primer

5`-CCGGAATTCATGGAGGGCAAAACCCGCACA-3`(EcoR I)

VP60 downstream primer

5`-ACGCGTCGACTTATCAGACATAAGAAAAGC-3`(Sal I)

CDSIII Primer

5′-ATTCTAGAGGCCGAGGCGGCCGACATG-d(T)30VN-3′)*

SMART III Oligo

5′-AAGCAGTGGTATCAACGCAGAGTGGCCATTATGGCCGGG-3′)

5′PCR Primer

5′-TTCCACCCAAAGCAGTGGTATCAACGCAGAGTGG-3′

3′PCR Primer

5′-GTATCGATGCCCACCCTCTAGAGGCCGAGGCGGCCGACA-3′

AD-5`primer

5`-CTATTCGATGATGAAGATACCCCCACCAAACCC-3`

AD-3`primer

5`-GTGAACTTGCGGGGTTTTTCAGTATCTAC...

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Abstract

The invention discloses a protein bonded with rabbit hemorrhagic disease virus (RHDV) VP60 protein and application thereof. The protein is characterized by comprising at least one sequence which is shown as SEQ ID NO: 1. In the invention, moesin (MSN) which interacts with rabbit hemorrhagic disease virus structural protein VP60 is tagged from a rabbit liver tissue complementary DNA (cDNA) librarythrough a yeast two-hybrid technology. Immunoprecipitation tests verify that the protein has the function of being bonded with virus particles in vitro; and the purified MSN protein can inhibit the agglutination effect of 8 hemagglutination units of virus antigen on human O-shaped erythrocytes. The protein and the rabbit hemorrhagic disease virus are incubated at room temperature; an anti-RHDV negative rabbit is inoculated through oral administration and nasal dripping, still survives after 8 days and is healthy-looking; and pathological anatomy and excisional biopsy show that each visceral organ is normal. The rabbit which is infected by the virus dies in 3 days. Thus, the protein can be used for preparation of medicaments for treating rabbit hemorrhagic disease.

Description

technical field [0001] The invention relates to a protein capable of interacting with rabbit hemorrhagic disease virus coat protein VP60 and its application, in particular to a moesin MSN and its application in the preparation of drugs for treating rabbit hemorrhagic disease, belonging to the field of biomedicine. Background technique [0002] Rabbit Hemorrhagic Disease (RHD), commonly known as "Rabbit Plague", is an acute, severe, contagious infectious disease caused by Rabbit Hemorrhagic Disease Virus (RHDV), characterized by liver necrosis, The main features are edema of parenchymal organs, congestion and hemorrhagic changes in the respiratory system and multiple organs of the whole body, which usually cause a devastating blow to the rabbit industry. As the only coat protein of the virus, VP60 still maintains a high degree of homology after more than 20 years of strain prevalence, and it plays an important role in disease infection and host immunity. [0003] The rapid m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47C12N15/12C12N15/63C12N1/15C12N1/19C12N1/21A61K38/17A61K48/00A61P31/14
Inventor 刘家森曲连东郭东春姜骞林欢
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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