Supercharge Your Innovation With Domain-Expert AI Agents!

Recombinant baculovirus and application thereof in preparing Newcastle disease virus-like particles

A technology of recombinant baculovirus and Newcastle disease virus, which can be applied in the directions of viruses, viral peptides, antiviral agents, etc., can solve the problems of accelerating virus mutation, and achieve the effect of reducing production costs.

Active Publication Date: 2020-06-30
YEBIO BIOENG OF QINGDAO
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In my country, vaccination is mainly used to prevent and control the spread of the disease. Traditional vaccines use the complete pathogen as the antigen for vaccine production, which has certain safety risks.
Moreover, the use of whole viruses will cause selection pressure on the virus and accelerate the mutation of the virus

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Recombinant baculovirus and application thereof in preparing Newcastle disease virus-like particles
  • Recombinant baculovirus and application thereof in preparing Newcastle disease virus-like particles
  • Recombinant baculovirus and application thereof in preparing Newcastle disease virus-like particles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1, Newcastle disease virus HN protein, F protein

[0020] In 2018, the applicant isolated a strain of Newcastle disease virus from chickens in a chicken farm in Shandong. Add the disease material to the normal saline containing double antibody at a ratio of 1:5 (w / v), grind it to make a suspension, centrifuge at 1000r / min for 5 minutes, take the supernatant and inoculate the allantoic cavity of 10-11 day-old SPF chickens Embryos, 0.2ml per embryo, hatched at 36°C-37°C for 3-4 days, irradiated the embryos twice a day, took the dead chicken embryos after 24 hours, cooled them at 2-8°C for 8-16 hours, harvested the chicken embryo liquid, and carried out HA and HI tests. 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 was Newcastle disease virus, free of bacteria, mycoplasma and foreign virus contamination...

Embodiment 2

[0022] Example 2: Construction of recombinant baculoviruses expressing HN and F genes

[0023] 1. Cutting and optimization of HN and F genes

[0024] Cut off the transmembrane region of 45 amino acids in front of the N-terminal domain of the HN gene; add the start codon ATG to the N-terminus; optimize the codons according to the preference of insect cells, and the optimized nuclear The nucleotide sequence is SEQ ID NO:5, and the amino acid sequence is SEQ ID NO:6. Cut off the first 142 amino acids at the N-terminal of the F gene, add the start codon ATG; cut off the transmembrane region of the 53 amino acids at the C-terminal, and optimize the codon at the same time, the optimized nucleotide sequence is SEQ ID NO:7, the amino acid sequence is SEQ ID NO:8. The cut and optimized sequences and the uncut and optimized sequences were synthesized by Shanghai Bioengineering Co., Ltd. and linked into the cloning vector.

[0025] 2. Construction of HN positive plasmid

[0026] 2.1 ...

Embodiment 3

[0102] Example 3 SF9 cell transfection

[0103] (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.

[0104] (2) Add 2 μg of the recombinant DNA prepared in Example 2 to 100 μl of TNM-FH culture 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 minutes.

[0105] (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.

[0106] (4) Add 2 ml of 10% fetal bovine serum TNM-FH culture solution to each cell well.

[0107](5) Gently add the mixture of recombinant DNA and liposomes into each well of cells, mix gently, and culture statically ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a recombinant baculovirus and an application thereof in preparing Newcastle disease virus-like particles. The provided recombinant baculovirus comprises a nucleotide fragment for encoding a Newcastle disease virus HN and F protein; and the amino acid sequences of the HN and F protein are SEQ ID NO: 6, and SEQ ID NO: 8 respectively. The recombinant baculovirus constructed bythe invention is used for preparing virus-like particles of Newcastle disease virus in insect cells; and the prepared virus-like particles are used for preparing vaccines. According to the method, thecDNA full sequence of Newcastle disease virus HN and F protein is screened and analyzed, and the cDNA sequence which is relatively rich in epitope and capable of efficiently expressing in insect cells is selected. After the HN and F proteins are optimized and transformed, the virus-like particles are prepared by using an insect cell-baculovirus expression system, the yield of the virus protein issignificantly higher than that of SPF embryo culture, and the production cost is obviously reduced.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering vaccines, in particular to a recombinant baculovirus and its application in preparing Newcastle disease virus-like particles. Background technique [0002] Newcastle disease (ND) is an acute and highly contagious disease of poultry caused by Newcastle disease vlrus (NDV). Since it first appeared in Indonesia and Newtown, UK in 1926, Newcastle disease has had many pandemics around the world in the history of more than 90 years. Newcastle disease has brought huge economic losses to my country's poultry industry and seriously threatened the development of my country's poultry industry, so the prevention and treatment of Newcastle disease is very important. [0003] Newcastle disease virus belongs to the family Paramyxoviridae, the genus Paramyxovirus, and its nucleic acid is single-stranded RNA. It consists of a helical and symmetrically coiled nucleocapsid and an envelope. There are r...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N7/01A61K39/17A61P31/14
CPCC12N7/00C07K14/005A61K39/12A61P31/14C12N2710/14021C12N2760/18122C12N2760/18123A61K2039/5258C12N2760/18134
Inventor 郭伟伟李金积王宗升陈俭梅向银辉刘大卫
Owner YEBIO BIOENG OF QINGDAO
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com