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

A Knockout Mutant Strain of Streptococcus suis type 4 galr gene sh1510δgalr

A technology of Streptococcus suis and gene knockout, applied in the field of genetic engineering, can solve the problem that SS4 virulence factor has not been reported, and achieve the effect of simple and efficient construction

Active Publication Date: 2021-06-04
JIANGSU ACAD OF AGRI SCI
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, domestic and foreign research on S. suis mainly focuses on S. suis type 2, and there are few studies on SS4, which are limited to serotyping PCR identification. The research on SS4 virulence factors has not been reported yet.

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
  • A Knockout Mutant Strain of Streptococcus suis type 4 galr gene sh1510δgalr
  • A Knockout Mutant Strain of Streptococcus suis type 4 galr gene sh1510δgalr
  • A Knockout Mutant Strain of Streptococcus suis type 4 galr gene sh1510δgalr

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Construction of Mutant SH1510ΔGalR

[0026] 1) Primers

[0027] The details of the primer sequences designed in this experiment are listed in Table 1. L1 / L2, R1 / R2 respectively amplify the gene sequence of the upstream and downstream homology arms of the GalR coding gene; C1 / C2 amplifies the chloramphenicol resistance gene; I1 / I2 amplifies the ORF internal gene sequence of the GalR gene (for deletion strains); design a pair of primers O1 / O2 outside the upper and lower homology arms of the GalR gene, and amplify the total length of a gene sequence including the GalR gene, the upper and lower homology arms genes and the outside (for external detection) .

[0028] Table 1 Primer sequence list

[0029]

[0030] 2) Construction of the deletion strain SH1510ΔGalR

[0031] A. Construction of the gene deletion plasmid pSET4s::GalR as follows figure 1 As shown, primers L1 / L2, R1 / R2 and SH1510 DNA templates were used to amplify the upstream and downstream homolo...

Embodiment 2

[0042] Example 2 Gram staining

[0043] The cryopreserved parent strain SH1510 and deletion strain SH1510ΔGalR plates were streaked for recovery, and single colonies of the parent strain and deletion strain were picked and inoculated in THB broth, 200 r / min, shaking culture at 37°C until logarithmic growth phase, The parental strain SH1510 and the deletion strain SH1510ΔGalR were subjected to smear, Gram staining and microscopic examination, and the morphology of the two strains was observed and compared under the oil lens.

[0044] exist Figure 4 It can be seen that the two strains are round or oval, arranged in chains, and have similar lengths, without significant difference.

Embodiment 3

[0045] The comparison of different sugar utilization rates of embodiment 3

[0046] The basal medium was prepared according to Tang Yulong's method, and six bottles of basal medium were prepared for the test. Two bottles were used as a group, and 10mmol / L glucose, sucrose and D-galactose were added respectively. Resuscitate the bacteria, pick a single colony of the parental strain SH1510 and the deletion strain SH1510ΔGalR and inoculate them in THB broth, 200r / min, shake at 37°C until OD 600 =0.7, centrifuge at 12000r / min for 10min, discard the supernatant, wash the bacteria twice with sterilized physiological saline, and resuspend the bacteria in an equal volume. Add the bacterium solution of the parental strain and the deletion strain to the medium of the three groups at a ratio of 1%, and measure the OD every 1 h 600 value until bacterial OD 600 The value can only be stopped when it reaches a stable value, the test is repeated 3 times, and the hourly OD is calculated 600...

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 Streptococcus suis type 4 GalR gene knockout mutant strain SH1510ΔGalR. The mutant strain SH1510ΔGalR has been preserved in the China Center for Type Culture Collection on June 20, 2019, and the number is CCTCC NO: M2019477. The mutant SH1510ΔGalR is obtained by knocking out the transcription regulator GalR gene in the genome of the virulent Streptococcus suis type 4 strain SH1510 and replacing it with a chloramphenicol resistance gene. The gene knockout plasmid pSET4s::GalR containing the homologous recombination fragment was constructed, and the gene knockout plasmid pSET4s::GalR was electrotransformed into Streptococcus suis serotype 4 SH1510 to obtain it. The mutant strain of the invention lays the foundation for further research on the function of Streptococcus suis type 4 GalR protein and its pathogenic mechanism.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, in particular to a type 4 GalR gene knockout mutant strain SH1510ΔGalR of Streptococcus suis. Background technique [0002] Streptococcus suis is an important pathogenic bacterium in pigs, which can cause swine arthritis, sepsis, endocarditis, meningitis, pneumonia and other diseases, and can also lead to morbidity and death of livestock and veterinary practitioners. According to the bacterial capsule antigen, Streptococcus suis can be divided into 33 serotypes (types 1-31, 33 and 1 / 2), among which Streptococcus suis type 2 is recognized as the most pathogenic serotype. In addition, serotypes 3, 4, 5, 7, 8, 1 / 2 often appear in Asian countries; serotypes 3, 4, 8, 22, 1 / 2 are often isolated in Canada. Streptococcus suis type 4 (SS4) is also a pathogenic serotype that can cause disease and even death in humans and animals. From 1968 to 1984, 30 strains of Streptococcus suis that caused...

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): C12N1/21C12N15/90C12N15/74C12N15/65A61K39/09A61P31/04C12R1/46
CPCA61K39/092A61K2039/52A61K2039/552A61P31/04C07K14/315C12N15/65C12N15/746C12N15/902
Inventor 倪艳秀孙珂祝昊丹王丹丹周俊明吕立新余正玉何孔旺李彬温立斌张雪寒郭容利胡屹屹汪伟范宝超肖琦张碧成朱雪蛟周金柱
Owner JIANGSU ACAD 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