Light-emitting reporting system and application thereof to antibiotic discovering and screening

A reporting system and antibiotic technology, applied in the biological field, can solve problems such as antibiotic discovery and screening that have not yet been achieved, and achieve the effects of increasing probability, speeding up discovery, and huge social and economic benefits

Active Publication Date: 2019-09-17
SHANDONG UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After searching, there is no report on the discovery and screening of antibiotics using the luminescent reporter system constructed from the promoter fragments of the molecular chaperone genes clpX, groE, dnaJ, dnaK and the luxABCDE gene fragment connected to the molecular chaperone gene promoter.

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
  • Light-emitting reporting system and application thereof to antibiotic discovering and screening
  • Light-emitting reporting system and application thereof to antibiotic discovering and screening
  • Light-emitting reporting system and application thereof to antibiotic discovering and screening

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Experimental strains and plasmids:

[0026] Escherichia coli DH5α: used for competent preparation, obtained from China General Microorganism Culture Collection Center (CGMCC1.12873).

[0027] Escherichia coli DH10B: used for competent preparation, obtained from ThermoFisher Scientific (Cat. No. 18297010)

[0028] Staphylococcus aureus RN4220: used for competent preparation and luminescence detection, acquired from BEI Resources.

[0029] pMD19-T: Purchased from Dalian Bao Biological Company (TaKaRa)

[0030] pAmilux: from the University of British Columbia, Canada (Document: Mesak LR, Yim G, Davies J, Improved lux reporters for use in Staphylococcus aureus, Plasmid, 2009, 61(3): 182-187.)

[0031] The medium and its preparation required for the experiment:

[0032] LB medium: For each liter of medium, tryptone (10g), sodium chloride (10g), and yeast extract (5g) should be added to 950ml of deionized water, and stirred with a magnetic stirrer until dissolved. Add 5 m...

Embodiment 2

[0146] Example 2 The response of the luminescent reporter system to antibiotics

[0147] Experimental strains: The strains used in the experiment are four lux reporter strains of Staphylococcus aureus: S1, S2, S3, S4, and the strain S0 containing pAmilux empty plasmid.

[0148] Media required for experiment and its preparation

[0149] 1. NYE agar medium

[0150] 2. 0.7% agar: Weigh 0.7g of agar with a balance and add it to the Erlenmeyer flask, then add 100ml of deionized water, and sterilize under high pressure steam at 121°C for 20 minutes for later use.

[0151] Antibiotics required for experiments and their preparation

[0152] 1. Daptomycin (DAP) solution: Dissolve 30 mg of daptomycin in 1 ml of ultrapure water to make the final concentration 30 mg / ml, and filter through a 0.22 μm membrane to sterilize;

[0153] 2. Polymyxin B (PMB) solution: Dissolve 100mg of polymyxin B in 1ml of ultrapure water to a final concentration of 100mg / ml, and filter through a 0.22μm membr...

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
diameteraaaaaaaaaa
Login to view more

Abstract

The invention discloses a light-emitting reporting system. The light-emitting reporting system is constructed from a promoter fragment of molecular chaperone genes clpX, groE, dnaJ and dnaK, and a luxABCDE gene fragment connected with molecular chaperone gene promoters. The invention further discloses an application of the light-emitting reporting system to antibiotic recovering and screening. Compared with an existing antibiotic screening technique, the light-emitting reporting system disclosed by the invention has the advantages that the existence of antibiotics in an solution can be identified in a quick and high-flux manner, the discovering speed of new antibiotics can be increased, quick discovering and screening of the antibiotics can be realized, and the light-emitting reporting system has great application prospects.

Description

technical field [0001] The invention relates to a luminescent reporter system and its application in discovery and screening of antibiotics. Belongs to the field of biotechnology. Background technique [0002] Bacterial antibiotic resistance due to the large-scale use of antibiotics is a major threat to human health. According to statistics, there are currently 700,000 deaths worldwide due to bacterial resistance to antibiotics. If the current phenomenon of antibiotic resistance is not suppressed, by 2050, 10 million people will die due to antibiotic resistance worldwide, surpassing tumors to become the number one cause of death for human beings. To suppress the phenomenon of antibiotic resistance, on the one hand, people need to conduct further research on the mechanism of drug resistance to suppress or limit the occurrence, spread, and level of drug resistance, and on the other hand, it is also necessary to continuously develop new antibiotics. This requires rapid and e...

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): C12N15/63C12N15/65C12Q1/18
CPCC12N15/63C12N15/65C12Q1/18
Inventor 徐海王明钰
Owner SHANDONG 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