Construction method and application of novel goose parvovirus SD strain full-length infectious clone causing duck short beak and dwarf syndrome

A goose parvovirus and infectious cloning technology, applied in the field of viruses, can solve problems such as no research and reports, and achieve the effect of concise construction steps and guaranteed integrity

Active Publication Date: 2022-05-13
HEBEI AGRICULTURAL UNIV.
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The establishment of a reverse genetic operating system for the new goose parvovirus can be used as a basis for research on the virulence and cross-species transmission mechanism of the new goose parvovirus, and it can also provide a useful platform for the research of related vaccines, but at present There are no related studies and reports

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
  • Construction method and application of novel goose parvovirus SD strain full-length infectious clone causing duck short beak and dwarf syndrome
  • Construction method and application of novel goose parvovirus SD strain full-length infectious clone causing duck short beak and dwarf syndrome
  • Construction method and application of novel goose parvovirus SD strain full-length infectious clone causing duck short beak and dwarf syndrome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Construction of the full-length infectious clone of the new goose parvovirus SD strain

[0044] The schematic diagram of the construction strategy of the full-length infectious clone of the new goose parvovirus SD strain is as follows figure 1 shown. The specific process is as follows:

[0045] 1.1 Virus strains and duck embryos

[0046] NGPV SD was isolated and preserved by the Veterinary Biological Products Laboratory of Hebei Agricultural University, and the measured whole genome sequence was uploaded to NCBI (GenBank serial number: KY511124). The fifth generation virus was used in this study.

[0047] 1.2 Construction of intermediate plasmid pSK-XBSE

[0048] NGPV SD genome structure such as figure 2 As shown, 1-189bp is marked as A fragment, 190-585bp is marked as B fragment, 586-4470bp is marked as C fragment, 4471-4868bp is marked as D fragment, and 4869-5053bp is marked as E fragment. The Sph Ⅰ restriction site was selected as the intermediate cl...

Embodiment 2

[0079] Example 2: Rescue of novel goose parvovirus SD strain infectious DNA

[0080] 2.1 Transfection

[0081] Using Lipofectamine 2000 as the transfection reagent, the plasmid and the transfection reagent were mixed at a ratio of 1:2.5 (μg:μL). Take 1mL of Opti-DMEM medium to dilute the plasmid (the amount of plasmid is 20μg), and mix gently; after gently shaking Lipofectamine2000, take 50μL and 950μL of Opti-DMEM medium, mix well, and incubate at room temperature for 5 minutes; The plasmid was mixed with the diluted Lipofectamine 2000, mixed gently, and left at room temperature for 20 minutes; the transfection mixture was passed through the chorioallantoic cavity and the allantoic cavity at 0.3 mL / piece, and the amount of plasmid inoculated per duck embryo was 3.0 μg. The allantoic fluid was harvested 120 hours after inoculation, and the duck embryos were inoculated with 0.3 mL / duck embryos for subculture through the same way. The allantoic fluid was harvested after 120 hou...

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 construction method and application of a novel goose parvovirus SD strain full-length infectious clone causing duck short beak and dwarf syndrome, and belongs to the technical field of viruses. The method comprises the following steps: cloning a complete genome of a novel goose parvovirus SD strain to a plasmid pBluescript II SK through a segmented amplification method to obtain a recombinant plasmid; meanwhile, the NcoI site in the genome is mutated by utilizing an overlapping PCR technology and is used as a genetic marker for identifying the wild virus and the rescue virus; mixing the recombinant plasmid with a transfection reagent, and inoculating an SPF duck embryo through a yolk sac and allantoic cavity way to obtain a rescued virus. The novel goose parvovirus reverse genetic operating system established by the invention can be used for research in the directions of novel goose parvovirus virulence analysis, cross-species transmission mechanism and the like, and also lays a foundation for research on novel vaccines of duck short beak and dwarf syndrome.

Description

technical field [0001] The invention belongs to the technical field of viruses, and in particular relates to a construction method and application of a full-length infectious clone of a novel goose parvovirus SD strain that causes duck short beak and dwarfism syndrome. Background technique [0002] Since 2015, commercial meat ducks (Cherry Valley Duck, Peking Duck, etc.) in Jiangsu, Shandong, Hebei and other regions of my country have experienced unexplained developmental delays, atrophy of the upper and lower beaks, protruding tongues, swelling, and downward bending. The characteristic disease is commonly known as duck "short beak and dwarfism syndrome" (Short beak and dwarfism syndrome, SBDS). The incidence rate of the disease is 10% to 30%, and it can reach more than 50% in severe cases. The weight of sick ducks is 20% to 30% lower than that of healthy ducks, and in severe cases it is only 50% of normal weight. The feed-to-meat ratio of infected ducks increased significan...

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): C12N15/11C12N15/70C12N15/66C12N15/85C12N7/00C12Q1/70C12Q1/686A61K39/23A61P31/20C12R1/93
CPCC12N15/70C12N15/85C12N15/66C12N7/00C12Q1/701C12Q1/686A61K39/12A61P31/20C12N2750/14152C12N2750/14121C12N2750/14134A61K2039/53A61K2039/525C12Q2521/301Y02A50/30
Inventor 袁万哲郝雪飘赵款雷白时薛拥志张武超
Owner HEBEI AGRICULTURAL 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