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Virus-like particles of duck derived goose parvoviruses and preparation method of virus-like particles

A virus-like, duck-derived goose technology, applied in biochemical equipment and methods, antiviral agents, viruses/phages, etc., can solve problems such as differences in parvoviruses, improve antibody levels, and prevent gosling plague virus in muscovy ducklings The effect of infection

Pending Publication Date: 2019-06-28
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disease first appeared in China and many European countries in the middle and late 1960s. It was not called goose parvovirus infection until 1978. The isolated parvoviruses were significantly different

Method used

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  • Virus-like particles of duck derived goose parvoviruses and preparation method of virus-like particles
  • Virus-like particles of duck derived goose parvoviruses and preparation method of virus-like particles
  • Virus-like particles of duck derived goose parvoviruses and preparation method of virus-like particles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1, screening of Muscovy duck parvovirus VP2 protein

[0018] In 2015, typical symptoms of muscovy duck parvovirus disease appeared in young muscovy ducks in a certain duck farm. In order to isolate the pathogen, the liver, spleen and pancreas of the dying duck were aseptically collected, homogenized with sterile saline to make a suspension, centrifuged to collect the supernatant bacteria, and then inoculated 11-day-old muscovy duck embryos through the allantoic cavity, and hatched for 168 After 24 hours, the allantoic fluid and embryo body tissue of the dead embryos were collected, homogenized, frozen and thawed repeatedly, and the supernatant was frozen for storage. After the harvested virus liquid was purified, the virus content, immunogenicity, specificity and purity were analyzed and tested for virus characteristics. The results showed that the isolated virus strain had a specific reaction with Muscovy duck parvovirus, and there were no bacteria and mycop...

Embodiment 2

[0021] Embodiment 2, the construction of the recombinant baculovirus expressing VP2 gene

[0022] 2.1 Cutting and optimization of VP2 gene

[0023] Cut the parvo_coat domain of the vp2 gene, cut off the N-terminal 70aa, cut off the C-terminal 31aa, and at the same time mutate the 336th V to P, so as to help the protein to better fold into a tertiary structure. Its antigenic sites are better exposed. The amino acid sequence after optimization and modification is SEQ ID NO: 3, which can well express the antigenic site and can be automatically assembled into VLPS. The selected gene was codon-optimized, and the nucleotide sequence of the optimized gene is SEQ ID NO:4.

[0024] 2.2 Enzyme digestion reaction

[0025] 2.2.1 Mark the 1.5mL EP tube to be used, and add and mix the sample in the 1.5mL EP tube according to the following table: the reaction system is 50 μL, and the sample addition is shown in the table below:

[0026]

[0027] 2.2.2 Place the 1.5mL EP tube in step 2...

Embodiment 3

[0094] Embodiment 3: the packing of baculovirus

[0095] (1) Preparation: UV sterilization in a biosafety cabinet for 30 minutes; TNM-FH culture solution was placed in a 27°C water bath and preheated to 27°C.

[0096] (2) Add 2 μg of recombinant DNA to 100 μl of TNM-FH medium without serum and double antibody, and mix well. Add 9 μl Cellfectin Reagent to 100 μl TNM-FH medium without serum and double antibody, and mix well. The liposomes were mixed with the recombinant DNA and allowed to stand at room temperature for 40 min.

[0097] (3) Take out the 6-well plate cells from the incubator at 27°C, discard the supernatant medium, wash the cells three times with pre-warmed TNM-FH culture medium, and discard the TNM-FH culture medium.

[0098] (4) Add 2 ml of 10% fetal bovine serum TNM-FH culture solution to each cell well.

[0099] (5) Gently add the mixture of recombinant DNA and liposomes into each well of cells, mix gently, and culture statically at 27°C for 5-6 hours.

[0...

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Abstract

The invention provides virus-like particles of duck derived goose parvoviruses. A recombinant baculovirus used for preparing the virus-like particles contains a nucleotide fragment encoding a VP2 protein; the amino acid sequence of the VP2 protein is SEQ ID NO:3; the sequence of the nucleotide fragment of an encoding gene is SEQ ID NO:4. The prepared virus-like particles are used for preparing a vaccine. The prepared vaccine has the advantages that after the vaccine immunizes the cairna moschata, the antibody level of the cairna moschata can be improved, the antibody level of parent sources ofoffspring of the cairna moschata is ensured, and the gosling plague virus infection of young cairna moschata caused by the cairna moschata derived gosling plague viruses is prevented.

Description

technical field [0001] The invention belongs to the technical field of poultry vaccines, and in particular relates to a virus-like particle of duck-derived goose parvovirus and a preparation method thereof. Background technique [0002] Goose parvovirus disease, also known as Derzsy's disease, is commonly known as goose flu, goose or gosling plague, goose hepatitis, goose enteritis, etc. It is a highly contagious disease that affects goslings and muscovy ducks. According to the age of infected goslings and ducklings, the disease can be manifested as acute, subacute and chronic. The acute type can cause 100% death of goslings within 10 days of age. Many countries have also reported a completely different antigenic Muscovy duck parvovirus infection with a mortality rate as high as 80%. The disease mainly occurs in 1 to 3-week-old gooses and young Muscovy ducks, especially those about 1 week old are more susceptible, and geese or Muscovy ducks over 4 weeks old rarely show cli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C07K14/015A61K39/23A61P31/14
CPCY02A50/30
Inventor 郭伟伟向银辉陈俭梅王丽萍邹敏范根成杜元钊
Owner YEBIO BIOENG OF QINGDAO
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