Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Novel coronavirus pneumonia paramyxovirus vaccine strain and construction method thereof

A technology for coronaviruses and paramyxoviruses, applied in the direction of viruses, viral peptides, antiviral agents, etc., can solve problems such as low production and failure to meet market needs

Active Publication Date: 2021-09-10
QINGDAO HARWARS BIOLOGY GRP LTD
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although vaccines are available at present, the output is relatively low, which is far from meeting the needs of the market

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
  • Novel coronavirus pneumonia paramyxovirus vaccine strain and construction method thereof
  • Novel coronavirus pneumonia paramyxovirus vaccine strain and construction method thereof
  • Novel coronavirus pneumonia paramyxovirus vaccine strain and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1 Construction method of novel coronavirus pneumonia paramyxovirus vaccine strain

[0018] (1) Establishment of cell lines expressing T7 RNA polymerase

[0019] Take the BL21 strain and shake the bacteria to extract the plasmid. Design primers (the underline indicates restriction sites):

[0020] T7-F: 5'-CTG CTCGAG CCACCATGAACACGATTAACATCGCTAAGAACGAC-3' (SEQ ID NO: 1);

[0021] T7-R: 5'-CTG TCTAGA TTACGCGAACGCGAAGTCCGACTCTAAGATGT-3' (SEQ ID NO: 2);

[0022] The enzyme cutting sites are: upstream Xho I, downstream Xba I (marked by underline); amplify with the extracted BL21 plasmid as a template, and recover the target fragment, about 2.6kb. The PCI-neo vector and the recovered target fragment were subjected to double enzyme digestion, and the positive fragment was glued back, ligated and transformed with T4 ligase, and the plasmid was identified by PCR and enzyme digestion after small extraction, and the positive plasmid was selected and sent to Sango...

Embodiment 2

[0064] Example 2 Biological activity identification of novel coronavirus pneumonia paramyxovirus vaccine strain

[0065] Western Blot test was used to detect the expression of S1 protein: the supernatant of cells transfected with the pBR322-CDV-NPF-S1 plasmid constructed in Example 1 was used as a sample group, and the supernatant of cells transfected with an empty vector was used as a control group for Western Blot detection. Electrophoresis was performed on the sample group, control group and protein marker, and the electrophoresis was stopped when the bromophenol blue dye was at the end of the gel, and the membrane was transferred, and the temperature was kept low during the transfer process. After transfer, the membrane was blocked with skimmed milk powder for 1 hour. Add an appropriate amount of primary antibody that can bind to the receptor region of the new crown S1 protein and incubate at 37°C for 1 hour, wash 4 times with TBST, add a fluorescently labeled secondary a...

Embodiment 3

[0067] Embodiment 3 novel coronavirus pneumonia paramyxovirus vaccine strain animal immunogenicity

[0068] The new coronavirus pneumonia paramyxovirus vaccine strain prepared by the present invention was used to immunize rabbits, and each rabbit was immunized by eye drops and nasal drops for 10 days. 4 A total of 30 rats were immunized with the virus liquid at a dose of EID50; at the same time, 10 rats were set as a normal saline control group; they were raised in different isolators respectively. After 19 days, blood was collected from the immunized animals, the blood was centrifuged at 1500r for 5 minutes, and serum was taken for antibody detection.

[0069] The animal immunogenicity test method using the Beagle dog as a model is the same as above.

[0070] Experimental results: Antibodies could be detected in the serum of animals treated with the novel coronavirus pneumonia paramyxovirus vaccine strain prepared by the present invention, and no antibodies were found in the...

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 discloses a novel coronavirus pneumonia paramyxovirus vaccine strain and a construction method thereof, belonging to the technical field of virus vaccine strain construction. The construction method of the novel coronavirus pneumonia paramyxovirus vaccine strain of the present invention is to combine N, F genes of Paramyxoviridae VII type Newcastle disease virus with P, M, H, The L gene is recombined, and the new coronavirus S1 gene is inserted between the P gene and the M gene of the recombinant virus to obtain the whole genome of the new coronavirus pneumonia paramyxovirus; it is constructed by reverse genetic operation. The novel coronavirus pneumonia paramyxovirus vaccine strain constructed by the invention can stably and efficiently express the new coronavirus S1 protein, and can induce the body to produce antibodies after being injected into animals. Moreover, experiments can be carried out in poultry and dogs, which can save time and reduce costs, and at the same time, because of its large output, it is more conducive to actual production.

Description

technical field [0001] The invention belongs to the technical field of virus vaccine strain construction, and in particular relates to a novel coronavirus pneumonia paramyxovirus vaccine strain and a construction method thereof. Background technique [0002] Since 2019, the novel coronavirus has been rampant and has spread widely around the world, causing a large number of casualties. Although there are vaccines coming out at present, the output is relatively low, which is far from meeting the needs of the market. Current studies have shown that the new crown S protein plays an important role in virus infection and the induction of neutralizing antibodies. Contents of the invention [0003] The object of the present invention is to provide a novel coronavirus pneumonia paramyxovirus vaccine strain and its construction method. [0004] In order to achieve the above object, the present invention adopts following technical scheme: [0005] A method for constructing a novel...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/01C12N15/50A61K39/215A61P31/14A61P11/00C12R1/93
CPCC12N7/00C07K14/005A61K39/12A61P31/14A61P11/00C12N2770/20022C12N2770/20034C12N2760/18121C12N2760/18122C12N2760/18144C12N2760/18421C12N2760/18422C12N2760/18444A61K2039/543A61K2039/575A61K39/215
Inventor 李明义刘刚衣春霖马凤龙王宗科夏玉洁崔晓辰杨珍常宏赏赵冰冰孙艳红
Owner QINGDAO HARWARS BIOLOGY GRP LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com