Microbial cell sensor for detecting toluene organic pollutants

A microbial sensor, biosensor technology, applied in the directions of microorganisms, microorganism-based methods, and microbial determination/inspection, etc., can solve the problems of inability to realize on-site detection and expensive complete sets.

Inactive Publication Date: 2011-04-20
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its accuracy and sensitivity are very high, but it requires a complete set of expensive precision analytical instruments and specialized laboratories, and this method cannot achieve on-site detection in some pollutant detection

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: the first step: inoculate single bacterium colony (carrier cell blank is inoculated simultaneously) in 50ml Erlenmeyer flask; Ampicillin final concentration is 100 μ g / ml; 37 spend the night culture

[0022] The second step: 5ml of LB medium is added with a set concentration of inducer (such as xylene concentration of 500 μm);

[0023] Step 3: Bacteria cultured to OD600=1.2;

[0024] Step 4: Dilute to OD600=0.4, take 5ml of the diluted culture solution and add it to the culture solution prepared in the second step; induce at 23 degrees;

[0025] Step 5: For bacteria, mix 40 μl of untransformed cells (vector cells) with 50 μl of transformed culture. (same cell concentration) add 10 μl 1M K 2 HPO 4 (pH 7.8), 20 mM EDTA (one solution). The mixture was quickly frozen (freeze-thawed) in a -70°C refrigerator (10min), and then the cells were transferred to room temperature and placed in a room temperature water bath (23°C or placed at room temperature for 30 ...

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PUM

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Abstract

The invention discloses a microbial cell sensor for detecting toluene organic pollutants and a detection method thereof, and relates to a microbial cell sensor for detecting toluene organic pollutants, which is constructed by using a regulatory sequence of pseudomonas putida degradation genes and a plasmid sequence of regulatory protein genes and commercial report genes, and a detection method thereof. The bacterial host is Escherichia coli, and is used for detecting toluene organic matters. The lowest detection concentration on the toluene is 40 micromoles per liter, and the highest detection concentration on the toluene is 1,000 micromoles per liter.

Description

technical field [0001] The invention relates to a construction method of a microbial cell sensor for detecting toluene organic pollutants and a detection method thereof. The bacterium is Escherichia coli. The biosorbent is used to detect toluene-like organic pollutants in the environment. Background technique [0002] In recent years, with the acceleration of industrialization and agricultural modernization, a large number of products such as synthetic substances and chemical pesticides have been used, and humans have continuously discharged a large amount of pollutants into the environment, which have threatened human health and even life. The widespread use of toluene organic compounds has seriously polluted the environment. Toluene-like organic matter has been listed as a blacklist for polluting water bodies and soil, and must be tested and controlled. [0003] At present, there are two main methods for monitoring and detecting pollutants: one is physical and chemical ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12N1/21C12N15/70C12R1/19
Inventor 庄国强于清呼庆
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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