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

dcstamp gene and its application

A gene and transgenic chicken technology, applied in the fields of molecular biology and genetic engineering, can solve the problem of breaking the balance between waterfowl and influenza virus, and achieve the effect of inhibiting replication

Inactive Publication Date: 2019-07-19
CHINA AGRI UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the continuous evolution of influenza viruses, the balance between waterfowl and influenza viruses is broken

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
  • dcstamp gene and its application
  • dcstamp gene and its application
  • dcstamp gene and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] A new anti-AIV infection gene DCSTAMP was discovered by analyzing the transcriptome data.

[0033] Waterfowl, including domestic ducks, are the natural hosts of H5N1 subtype influenza virus, and some H5N1 subtype viruses do not show obvious clinical symptoms after infection. As the virus continues to evolve, strains with high pathogenicity to waterfowl also appear. In this study, two strains of H5N1 subtypes (highly pathogenic A / duck / Hubei / 49 / 05 (DK / 49) and low pathogenic A / goose / Hubei / 65 / 05 (GS / 65) were selected. ) H5N1 virus), infecting 4-week-old Shaoxing ducklings. Through the method of high-throughput sequencing, the gene expression profiles of spleen, brain and lung tissues of ducks infected with DK / 49 or GS / 65H5N1 influenza virus 1 day, 2 days and 3 days, and control ducks were respectively constructed, and the genes involved in the body's anti-inflammatory activity were identified. The immune gene related to influenza virus provides a new method for the treatm...

Embodiment 2

[0036] Obtaining the Full-length Coding Region Sequence of Duck DCSTAMP Gene Using Molecular Biology Experimental Methods

[0037] Referring to the duck genome reference sequence on the Ensemble website, and according to the transcriptome splicing sequence, design the duck DCSTAMP gene full-length CDS region cloning primer TF / TR, the sequence is as follows:

[0038] TF: 5'-ATGCAAGCACTTGTCTCAACAGCCC-3',

[0039] TR: 5'-TCCAAAACAC TGCTGACCAT TAGCC-3'.

[0040] Using the cDNA of duck spleen tissue as a template, NEB company's Q5 high-fidelity polymerase was used for PCR amplification, and the amplified product was recovered by gel and ligated to T carrier for sequencing. After sequence comparison analysis, it was found that the full-length coding region of the duck DCSTAMP gene is 1431bp (SEQ ID NO.1), encoding a total of 476 amino acids (SEQ ID NO.2).

[0041] The inventors successfully cloned the full-length coding region sequence of the duck DCSTAMP gene.

Embodiment 3

[0043]Transient overexpression of DCSTAMP gene in cells by cytology experiment method

[0044] 1. Construction of duck DCSTAMP gene overexpression vector

[0045] According to the coding region sequence of duck DCSTAMP gene, its eukaryotic expression vector primer eTF / eTR was designed, and the upstream and downstream primers were respectively introduced into Mlu I and Pme I restriction sites (underlined); A kozak (marked in bold) sequence was introduced before ATG, and a flag tag (marked in bold) was introduced at the C-terminus of the target gene. The primer sequences are as follows:

[0046] eTF:

[0047] 5'-CG ACGCGT GCCACCATGCAAGCACTTGTCTCAACAGCCCAGAATGC-3',

[0048] eTR:

[0049] 5'-GG GTTTAAAC CTACTTATCGTCGTCATCCTTGTAATCCACCACATTGTCATTTACCATTGTC-3'.

[0050] The CDS sequence of the amplified DCSTAMP gene (as shown in the sequence table CDS) was introduced into the original vector PiggyBac-X gene (constructed by the inventor's laboratory in the early stage, and t...

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 DCSTAMP gene of ducks and an amino acid sequence of codes of the DCSTAMP gene. The DCSTAMP gene of the ducks and the amino acid sequence have the advantages that the DCSTAMP gene of the ducks can be used for inhibiting influenza viruses, particularly, copying of AIV (avian influenza viruses) can be obviously inhibited, accordingly, the DCSTAMP gene of the ducks can be deeply functionally studied, and anti-AIV key protein structural domains or amino acid can be determined; a novel way can be provided for utilizing gene editing methods of transgenic technologies and the like for breeding excellent varieties of transgenic agricultural animals capable of resisting different types of viruses such as the AIV.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology and genetic engineering, and in particular relates to a DCSTAMP gene and its application. Background technique [0002] Influenza viruses are negative-strand RNA viruses containing eight RNA genome segments. Highly pathogenic avian influenza (Highly Pathogenic Avian Influenza, HPAI) is a severe infectious disease mainly in poultry caused by influenza A virus of the Orthomyxoviridae family. [0003] Waterfowl, including domestic ducks, are the natural hosts of influenza virus, and all subtypes of different combinations of 16 HA and 9 NA could be isolated in waterfowl. Influenza virus has always maintained the characteristics of low pathogenicity to waterfowl. Waterfowl infected with the virus will not get sick, but they can shed the virus to the outside world. Certain strains with low pathogenicity to waterfowl are highly pathogenic to chickens or other hosts. However, with the contin...

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): C12N15/12C07K14/465C12N15/85C12N5/10A01K67/027
CPCC07K14/465
Inventor 李宁黄银花荣恩光王小雪
Owner CHINA AGRI UNIV
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