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

Mycoplasma bovis Mbov_0274 gene mutant and application thereof

A technology of Mycoplasma bovis and mutant strains, applied in veterinary vaccines, bacterial antigen components, antibody medical components, etc., can solve the problems that hinder the development of new drugs and new vaccines, prevent the effective prevention and treatment of Mycoplasma bovis, and the pathogenic mechanism is unclear. And other issues

Active Publication Date: 2020-10-16
HUAZHONG AGRI UNIV
View PDF6 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the unclear pathogenic mechanism of M. bovis seriously hinders the development of new drugs and vaccines, and thus hinders the effective prevention and treatment of M. bovis disease.

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
  • Mycoplasma bovis Mbov_0274 gene mutant and application thereof
  • Mycoplasma bovis Mbov_0274 gene mutant and application thereof
  • Mycoplasma bovis Mbov_0274 gene mutant and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] Example 1: Construction of Mycoplasma Bovis Inserted Mutant Library

[0011] The applicant isolated a strain of Mycoplasma bovis HB0801 from the diseased lung tissue of sick cattle in June 2008, and named it Mycoplasma bovis HB0801, Mycoplasma bovis HB0801, and this strain has been disclosed in the patent literature of CN 102220263A.

[0012] The pMT85 plasmid was gifted by Dr. Eric Baranowski of the French Academy of Agricultural Sciences. It contains a mini-Tn4001 transposon (mini-Tn4001) into which the gentamicin resistance marker encoded by the aacA-aphD gene has been introduced. Located between the two inverted repeats (IR) at both ends of the transposable fragment, and the transposase gene (tnpA) is located outside the repeat, which can prevent transposition from happening again (Baranowski et al 2010).

[0013] To construct a mutant library with M.bovis HB0801 as the parent strain, the specific operation is: collect the M.bovis cultured to the late logarithmic stage, wa...

Embodiment 2

[0014] Example 2. Genome sequencing and identification of mutant strains of Mycoplasma bovis mutant library

[0015] Using the bacterial genome extraction kit (purchased from Bao Bioengineering Dalian Co., Ltd.), extract the total DNA of Mycoplasma bovis in the Mycoplasma bovis mutant library, sequence the junction of the Tn4001 transposon and the Mycoplasma bovis genome, and compare the sequencing results with Mycoplasma bovis The whole genome sequence of HB0801 was compared, and the results showed that the gene Mbov_0274 involved in the mutant strain T9.202 covered the transposon insert sequence, the size was 3438bp, located after the 315328 position of the genome, and after the 1664 position of the Mbov_0274 gene ( figure 1 ).

Embodiment 3

[0016] Example 3: Verification of the ability of T9.202 mutant to stimulate BoMac cells to express cytokines

[0017] Culture and counting of Mycoplasma bovis: Take the highly virulent Mycoplasma bovis strain HB0801, the weakly virulent Mycoplasma bovis strain MbovHB0801-150.2, and T9.202 to inoculate the PPLO liquid medium at a ratio of 1:1000, and place it at 37℃, 5% CO 2 After 36 hours of culture in the incubator to reach the logarithmic phase, the CFU count is carried out. The method is as follows: Dilute the cultured bacterial solution in 10-fold increments, and apply 10 μL of the bacterial solution with an appropriate dilution to the PPLO solid medium at 37°C. Inverted culture, 5% CO 2 After culturing in the incubator for 3 to 7 days, colony count is performed under a stereo microscope. The calculation formula of the colony number is: CFU / mL = number of colonies × dilution × 100.

[0018] Place BoMac 2×10 per hole 6 Cell density is seeded into 6-well cell plate, 37℃, 5% CO 2 C...

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 mycoplasma bovis Mbov_0274 gene mutant, and belongs to the technical field of animal borne disease prevention and treatment. The mutant is named as mycoplasma bovis T9.202, is preserved in China Center for Type Culture Collection with the preservation No. of CCTCC NO:M 2020084, and the nucleotide sequence of the Mbov_0274 gene is shown as SEQID NO:1. The capacity of a T9.202 strain for stimulating expression of IL-8 by BoMac cells is detected by qRT-PCR, results show that the capacity of the mycoplasma bovis bacterial strain T9.202 for inducing the expression of cytokine by the BoMac cells is reduced, and the mycoplasma bovis Mbov_0274 gene mutant is hopeful to development of important effects in the field of mycoplasma bovis immunization control.

Description

Technical field [0001] The invention belongs to the technical field of prevention and treatment of animal infectious diseases, and specifically relates to a Mbov_0274 gene mutant strain of Mycoplasma bovis. The mutant strain induces a reduced ability of macrophages to express IL-8 cytokines and has application potential in preparing vaccines for Mycoplasma bovis. Background technique [0002] Mycoplasma bovis (M.bovis) belongs to the Mollicutes (Mollicutes), Mycoplasmatles, Mycoplasma family, and Mycoplasma. It is an important pathogen that harms the development of cattle breeding and is widespread worldwide , Can infect cattle of any age, and its incidence is mostly related to transportation stress. In most cases, it is manifested as chronic pneumonia and arthritis. Lactating dairy cows mainly manifested with Mycoplasma bovis mastitis, and calves suffered from Mycoplasma bovis pneumonia and arthritis through milk infection. With the increase of clinical antibiotic resistance, t...

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): C12N1/20A61K39/02A61P31/04C12R1/35
CPCA61K39/0241A61K2039/552A61P31/04C12N1/205C12R2001/35
Inventor 郭爱珍赵刚伊赫萨努拉·希拉尼张慧陈颖钰胡长敏陈曦陈建国
Owner HUAZHONG 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