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

Water quality genotoxicity detection method based on semiconductor opening switch (SOS) effect of recombinant Escherichia coli

A technology for recombinant Escherichia coli and Escherichia coli, which is applied in the direction of material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of high price, insensitivity of chlorinated organic substances, and difficult access to recombinant bacteria system, and achieves simple and easy cultivation and operation , Improve the sensitivity of fluorescence detection and reduce the effect of detecting fluorescence background value

Active Publication Date: 2012-10-31
济南市供排水监测中心
View PDF3 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This system has the following problems in practical application: 1) It is not sensitive to chlorinated organic substances, which is not conducive to the detection of disinfection by-products in water bodies, especially in drinking water; Nearly 100 million people in the world have been infected, some of them have died, and there are microbial risks in the operation of the constructed Salmonella typhimurium system; 3) The application steps of the system are relatively cumbersome, and the detection of β-galactosidase activity requires the addition of substrates, Such as β-ONPG (o-nitrophenyl β-D-galactoside), the price is relatively expensive
4) Involving intellectual property rights, it is still difficult to obtain the recombinant bacterial system, and it is difficult to widely promote it in the domestic industry

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
  • Water quality genotoxicity detection method based on semiconductor opening switch (SOS) effect of recombinant Escherichia coli
  • Water quality genotoxicity detection method based on semiconductor opening switch (SOS) effect of recombinant Escherichia coli
  • Water quality genotoxicity detection method based on semiconductor opening switch (SOS) effect of recombinant Escherichia coli

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The application of the genetic toxicity biological detection method based on the SOS effect of recombinant E. coli in the detection of concentrated water samples (4L of water concentrated to 1mL) from a water plant, the steps are as follows:

[0043] (1) Resuscitation and activation of recombinant E. coli storage

[0044] Put 200μL of recombinant E. coli stock into 5mL modified LB liquid medium (tryptone 10g / L, yeast extract 2.5g / L, NaCl 10g / L). In addition, add glucose to account for 0.3% (W / V), add ampicillin to a final concentration of 50μg / mL, 30°C, and shake culture at 150rpm for 16-18 hours.

[0045] (2) Preparation before E. coli detection solution

[0046] Put the resuscitation solution into fresh modified LB liquid medium at a volume ratio of 1 / 100, shake culture at 35-37°C and 180 rpm, and cultivate until the OD (520nm) of the bacterial solution is 0.4 for about 2 hours.

[0047] (3) Contact with the tested sample

[0048] Add 100μL of the test sample to the prepared 1m...

Embodiment 2

[0062] The same method was used to detect the concentrated sample of the incoming water from the water plant in Example 1 (4L of water was concentrated to 1mL), and the ratio of fluorescence intensity was plotted against Cr 6+ Concentration standard curve, chromium ion concentration is used to express the water quality toxicity of the water sample. The difference is: when resuscitation, the culture temperature is 33℃, and the rotation speed is 180rpm; when preparing the detection solution, the rotation speed is 250rpm, and the culture is cultured to the OD (520nm) It is 0.3; when the water sample or chromium ion solution to be tested is in contact with the test solution, the amount added is 20% of the test solution volume, and the speed is 250rpm. After testing, the fluorescence intensity ratio of the water sample to be tested is 1.90, which is brought into the standard curve, and the genotoxicity of the water sample to be tested is equivalent to 0.0171 mg / L Cr 6+ The effect.

Embodiment 3

[0064] The same method was used to detect the concentrated sample of the incoming water from the water plant in Example 1 (4L of water was concentrated to 1mL), and the ratio of fluorescence intensity was plotted against Cr 6+ Concentration standard curve, the chromium ion concentration is used to express the water toxicity of the water sample, the difference is: the test solution after contact with the water sample or chromium ion solution is centrifuged at 10000rpm to separate the E. coli contact liquid for 2 minutes, and then broken by ultrasonic method The bacterial cells were broken by a program of 4 seconds, 4 seconds apart, and 15 times broken, and then the broken bacterial cells were centrifuged at a high speed of 14000 rpm for 8 minutes. After testing, the fluorescence intensity ratio of the water sample to be tested is 2.08, which is brought into the standard curve, and the genotoxicity of the water sample to be tested is equivalent to 0.0391mg / L Cr 6+ The effect.

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 water quality genotoxicity detection method based on a semiconductor opening switch (SOS) effect of recombinant Escherichia coli. The method comprises the following steps of: recovering, activating and culturing recombinant Escherichia coli storage to obtain Escherichia coli detection solution; contacting a water sample to be measured with the detection solution, and simultaneously setting a contrast; centrifuging, ultrasonically disintegrating and performing other post-treatment on the contacted detection solution, and detecting a fluorescence intensity ratio of thewater sample to be measured; measuring a fluorescence intensity ratio of the Cr6+ solution with different concentrations, which is contacted with the detection solution by using the same method; drawing a standard curve between the fluorescence intensity ratio and the Cr6+ concentration, and substituting the fluorescence intensity ratio of the water sample to represent the water quality toxicity of the water sample through the Cr6+ concentration. The method has no pathogenic risk, is easy and convenient to culture and operate, high in detection sensitivity and low in cost; the final result isrepresented by Cr6+ equivalent concentration, and can be compared with results of other biological detection methods; and the method is suitable for rapidly detecting genotoxicity of an environmentalsample.

Description

technical field [0001] The invention relates to the technical field of genotoxicity detection of environmental pollutants, in particular to a method for detecting genotoxicity of environmental pollutants by using recombinant Escherichia coli carrying a reporter gene. Background technique [0002] The genetic toxicity analysis methods of environmental pollutants are divided into long-term tests and short-term tests. Because long-term test methods are time-consuming, laborious, and expensive for long-term maintenance of experimental animals, they have been gradually replaced by fast and cheap short-term screening methods. Short-term testing uses cytogenetic indicators to screen chemical mutagens, and usually uses biological cells such as plants, aquatic organisms, mammals, and microorganisms to monitor environmental mutagens. Currently, bacterial reverse mutations, micronucleus technology, and sister chromatids have been developed. Exchange test, SOS reaction test, single-cell...

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): G01N21/64
Inventor 李力贾瑞宝孙韶华宋艳
Owner 济南市供排水监测中心
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