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

Recombinant adenovirus expressing mycoplasma hyopneumoniae P102 protein and applications thereof

A technology of mycoplasma hyopneumoniae and recombinant adenovirus, which is applied in the fields of application, virus/bacteriophage, and recombinant DNA technology, can solve problems such as the impact of inactivated vaccines and attenuated vaccines on popularization and application, and achieve low human pathogenicity, high safety, low probability effect

Inactive Publication Date: 2010-03-03
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the prevention and control of Mhp mainly adopts drug treatment, immunization and population purification. Vaccine immunization mainly uses inactivated vaccines and attenuated vaccines. However, due to various factors, the promotion and application of inactivated vaccines and attenuated vaccines are affected.

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 adenovirus expressing mycoplasma hyopneumoniae P102 protein and applications thereof
  • Recombinant adenovirus expressing mycoplasma hyopneumoniae P102 protein and applications thereof
  • Recombinant adenovirus expressing mycoplasma hyopneumoniae P102 protein and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1P102

[0039] The mutation of embodiment 1P102 gene

[0040] 1.1 Primer design and PCR amplification

[0041] The 5' end primer of primer A1 is provided with CCACC sequence, ATG promoter, Hind III site, and the 3' end primer is provided with TAA stop codon and Xho I site.

[0042] A1:

A2:

ATGGATCCAAACCTTTTTGGTTAATAA

CAGGTCGACATAATCAGAGAGGCTAAT

Primers for amplifying the A-segment gene

a1:

a2:

AAAAAATGGTCGGCAAAAACAGT

CCGACCATTTTTTAAAATTATCTTTAATT

Mutation at position 241-243

Primer

a3:

a4:

GCTCTTTCAAGTCGTGAAAACCAATTAC

TTCACGACTTGAAAGAGCTTTGGTTG

Mutation at position 610-612

Primer

a5:

a6:

TCTAAAATTCTAAAATCCTGGCTTGAAA

ATTTTAGAATTTTAGAAATTTCCTGATTTG

Mutation at position 877-879

Primer

B1:

B2:

CAGGAATTCTGGTATGGATCTCCGAATTCA

CAGGTCGACAAAAGTGCTTACCTCTGTT

Amplify B segment gene primers

C1:

C2:

CAGGGATCCGCAGAAGAAGCTAAG

...

Embodiment 2

[0051] Example 2 Construction and Expression of Mycoplasma Hyopneumoniae P102 Gene Recombinant Adenovirus

[0052] 1 Materials and methods

[0053] 1.1 Strains, plasmids, cells, gene fragments

[0054] Mycoplasma hyopneumoniae Yin-1 strain was isolated and identified in 2005 by our laboratory; P102 gene (mutated sequence SEQ ID NO: 1 obtained in Example 1); E.coliDH5α is a product of Novagen; adenovirus Expression vector system (AdEasy TM XLAdenoviral Vector System) including the shuttle vector pShuttle-CMV, electroporation competent cells BJ5183-AD-1, packaging cells AD-293 and super competent cells XL-10 were purchased from Stratagene.

[0055] 1.2 Main reagents

[0056] Tissue / cell DNA extraction kits, small plasmid extraction kits and gel recovery kits are all products of Shanghai Huashun Company; protein molecular quality standards, restriction endonucleases NotI, HindIII and Pyrobest TM DNA polymerase was produced by Fermentas MBI Company; restriction enzymes Pme I...

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

PropertyMeasurementUnit
Titeraaaaaaaaaa
Login to View More

Abstract

The invention discloses a recombinant adenovirus expressing mycoplasma hyopneumoniae P102 protein and applications thereof. The recombinant adenovirus is obtained by firstly carrying out mutation on gene of the mycoplasma hyopneumoniae P102 protein, cloning the whole genome (SEQ ID No:1) after the mutation onto a shuttle vector pShuttle-CMV, obtaining a recombinant shuttle vector, carrying out electro-transformation into competent cells containing an adenovirus backbone vector after linearization and carrying out homologous recombination in bacteria. The indirect immunofluorescence and the Western Blot identification prove that the recombinant adenovirus successfully expresses the P102 protein. The recombinant adenovirus is used for immunizing female Balb / c mice with 6-8 weeks by two waysof muscle injection and nasal dropping, and the recombinant virus can induce the mice to produce the specific humoral immune and the cellular immune response, thereby indicating that the recombinant adenovirus can be applied in the diagnosis, the prevention or the treatment of mycoplasma pneumoniae of swine.

Description

technical field [0001] The present invention relates to a rearranged cDNA sequence and a recombinant adenovirus containing the cDNA sequence, in particular to a cDNA sequence obtained by rearranging and optimizing the gene encoding the Mycoplasma hyopneumoniae P102 protein, and the present invention also relates to a recombinant adenovirus containing the cDNA The sequenced recombinant adenovirus and their application in the diagnosis, prevention or treatment of swine mycoplasma pneumonia belong to the field of prevention and treatment of swine mycoplasma pneumonia. Background technique [0002] Mycoplasma hyopneumoniae (Mycoplasma, hyopneumoniae, Mhp) is the main pathogen of swine mycoplasma pneumonia (MPS), which is also called porcine endemic pneumonia (EPP). . MPS is characterized by contact, highly contagious, chronic, high morbidity, and low mortality. The main manifestations are cough, dyspnea, and fleshy or marbled lesions in the lung tissue can be seen on dissection...

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): C12N15/31C12N15/63C12N7/01C07K14/30C12N15/70C12Q1/68G01N33/53A61K48/00A61K39/02A61P11/00A61P31/04C12Q1/70C12R1/35C12R1/19C12R1/93
Inventor 辛九庆李媛郭丹郭军陈超曹培丽张美晶董惠姜海芳
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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