Chimeric Newcastle disease virus vector H9 living vaccine candidate strain capable of overcoming influence of Newcastle disease maternal antibody in young chickens and construction method of candidate strain

A technology of Newcastle disease virus and maternal antibody is applied in the field of chimeric Newcastle disease virus vector H9 live vaccine candidate strain that overcomes the influence of Newcastle disease maternal antibody in chicks and its construction, and can solve problems such as restricting the application of NDV vector vaccine.

Inactive Publication Date: 2017-12-05
YANGZHOU UNIV
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This greatly limits the application of NDV vector vaccines in commercial chicken flocks
So far, there is still no H9 live vaccine with NDV as the carrier on 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
  • Chimeric Newcastle disease virus vector H9 living vaccine candidate strain capable of overcoming influence of Newcastle disease maternal antibody in young chickens and construction method of candidate strain
  • Chimeric Newcastle disease virus vector H9 living vaccine candidate strain capable of overcoming influence of Newcastle disease maternal antibody in young chickens and construction method of candidate strain
  • Chimeric Newcastle disease virus vector H9 living vaccine candidate strain capable of overcoming influence of Newcastle disease maternal antibody in young chickens and construction method of candidate strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Construction of recombinant plasmid pNDV / rAI4-T4FHN

[0034] Construction Step 1: Modification of Newcastle Disease Virus AI4 Genome

[0035] Biomaterial preparation:

[0036] The full-length expression vector pNDV / rAI4 of NDV AI4 strain (Hu Z, Hu S, Meng C, et al. Generation of a Genotype VII Newcastle Disease Virus Vaccine Candidate with High Yield in Embryonated Chicken Eggs. Avian Diseases 2011,55(3):1759- 1769) was constructed and preserved by the Key Open Laboratory of Livestock and Poultry Infectious Diseases of the Ministry of Agriculture of Yangzhou University.

[0037] pCR2.1 vector: purchased from Invitrogen; AMV reverse transcriptase, High fidelity DNApolymerase, T4DNA ligase, and Agarose Gel DNA Extraction Kit were purchased from Roche; transfection reagent SuperFect and plasmid extraction kit (QIAprep Spin MiniPrep Kit) were The products of QIAGEN; the rest of the conventional reagents were of domestic analytical grade.

[0038] Design mutati...

Embodiment 2

[0078] The construction model of recombinant plasmid pNDV / rAI4-T4FHN-H9 is as follows figure 1 .

[0079] Construction step 1: Construction of a full-length expression clone expressing HA protein of avian influenza H9N2 subtype strain using chimeric Newcastle disease virus AI4-T4FHN as a vector

[0080] Biomaterial preparation:

[0081] A / chicken / jiangsu / WJ57 / 2012 (WJ57), the H9N2 subtype epidemic strain of avian influenza virus, was isolated and preserved by the Key Open Laboratory of Livestock and Poultry Infectious Diseases, Ministry of Agriculture, Yangzhou University. The GenBank serial number is KJ000710.1

[0082] pCR2.1 vector: purchased from Invitrogen; Dephospharylation (BAP) Kit: purchased from Bao Biological Engineering (Dalian) Co., Ltd.; AMV reverse transcriptase, High fidelity DNA polymerase, T4 DNA ligase, AgaroseGel DNA Extraction Kit were purchased from Roche; Transfection reagent SuperFect and plasmid extraction kit (QIAprepSpin MiniPrep Kit) are product...

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 relates to a chimeric Newcastle disease virus vector H9 living vaccine candidate strain capable of overcoming influence of Newcastle disease maternal antibody in young chickens and a construction method of the candidate strain. Preservation number of the Newcastle disease vaccine strain is CGMCC No.13797. According to the construction method, by virtue of a reverse genetics manipulation platform which is established and is capable of achieving efficient replication of chimeric virus AI4-T4FHN with the presence of the Newcastle antibody, HA sequence of an avian influenza H9N2 strain is interpolated into an AI4-T4FHN strain genome whole-length transcription vector pNDV / rAI4-T4FHN, so that recombinant Newcastle disease virus genome whole-length cDNA cloning is achieved. The recombinant virus AI4-T4FHN-H9, which is obtained from transfection, has a relatively high propagation tiler on a chicken embryo, and stable expression of HA protein can be still guaranteed after continuous passage; and meanwhile, effective replication in a chicken body at a Newcastle disease high maternal antibody level can be implemented; therefore, the candidate strain is applicable to large-scale production of vaccines and is suitable for producing vaccines.

Description

technical field [0001] The invention relates to the application of reverse genetic technology to rescue a recombinant vaccine strain AI4-T4FHN-H9 expressing the HA protein of avian influenza H9 subtype virus by using chimeric Newcastle disease virus as a carrier, and is used for vaccine development. Background technique [0002] Reverse genetics manipulation technique (Reverse genetics manipulation technique) refers to the reverse transcription of RNA virus genomic RNA into cDNA, various in vitro artificial manipulations are performed on it at the DNA molecular level, and new viral genome cDNA and various auxiliary proteins are assembled. A research technique for RNA viruses [Neumann G, Whitt M A, Kawaoka Y. Adecade after the generation of a negative-sense RNA virus from cloned cDNA–what have we learned? Journal of General Virology, 2002, 83(11): 2635-2662.], also known as full-length infectious cDNA cloning technology, and often referred to as "virus rescue". Since the "re...

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 Applications(China)
IPC IPC(8): C12N7/01C12N15/85C12R1/93
CPCC12N7/00C12N15/85C12N2760/18021C12N2800/107
Inventor 刘秀梵刘晶晶胡顺林刘晓文王晓泉
Owner YANGZHOU UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap