Microbial cell sensor for detecting arsenic bioavailability degree

A technology of microbial cells and sensors, applied in the field of microbial cell sensors, can solve the problems of not reflecting the bioavailability of arsenic, expensive instruments, complicated operations, etc.

Inactive Publication Date: 2012-11-28
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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  • Abstract
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  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to solve the problems that the existing chemical detection method cannot reflect the bioavailability of arsenic and has complicated operation and expensive equipment, and utilizes the arsenic promoter sequence of the Escherichia coli arsenic resistance operon, the regulatory protein gene arsR and the commercial The luciferase reporter gene (luc) of the plasmid pEGMluc was constructed to construct a microbial cell sensor, thus providing a highly sensitive and low-cost method for the detection of arsenic bioavailability

Method used

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  • Microbial cell sensor for detecting arsenic bioavailability degree
  • Microbial cell sensor for detecting arsenic bioavailability degree
  • Microbial cell sensor for detecting arsenic bioavailability degree

Examples

Experimental program
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Embodiment 1

[0023] Step 1: Inoculate a single colony of sensor cells in a 50mL Erlenmeyer flask, add ampicillin to a final concentration of 100μg / mL, 37°C, 200r min -1 Overnight culture;

[0024] Step 2: Take 0.5mL of the above bacterial solution into 14.5mL of fresh LB medium, 37°C, 200r min -1 Grow to OD 600 = 1.2;

[0025] Step 3: Dilute the bacterial solution to OD with fresh LB medium 600 =0.4;

[0026] Step 4: Take 50 μL of the diluted bacterial solution and mix them with the arsenic standard solution and the sample to be tested in equal volumes, and let stand at 30°C for induction;

[0027] Step 5: Mix 40 μL empty vector cells with 50 μL induction medium, add 10 μL 1M K 2 HPO 4 (pH7.8) and 20mM EDTA lysis buffer, quickly freeze the mixture at -70°C for 10 minutes, then bathe the cells in water at 23°C for 3 minutes, and finally add 300 μL of freshly prepared lysis mixture (see appendix), mix well and incubate at room temperature for 10 minutes;

[0028] Step 6: Add 20 μL of ...

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Abstract

The present invention relates to a microbial cell sensor for detecting arsenic bioavailability degree, which is suitable for detecting bioavailability degree of arsenic in water and soil. The microbial cell sensor comprises: Escherichia coli as a host cell carrying recombinant plasmid. The recombinant plasmid is pUC18 plasmid containing arsenic resistant system ars promoter, arsenic resistant system controlling gene arsR, luciferase gene luc and rrnb terminator tandem sequence. The response time of the sensor on the arsenic is 30 minutes, the lowest detection concentration of the arsenic is 0.01 micromole/L, the highest detection concentration is 100 micromole/L. The microbial cell sensor has features of high sensitivity, fast response speed, low cost, and simple operation, and can be widely used in detecting the arsenic polluting the environment and evaluating the risk.

Description

technical field [0001] The invention relates to the construction and use of a microbial cell sensor for detecting the bioavailability of arsenic in water and soil. Background technique [0002] With the development of society and economy, the pollution situation of arsenic and its compounds has become increasingly severe, and has been assessed by WHO as one of the most serious heavy metal pollutants in water bodies. Arsenic and its compounds can cause comprehensive harm to the human cardiovascular system, nervous system and respiratory system. Reasonable and accurate monitoring of arsenic-polluted environment is an important prerequisite for pollution prevention and control. [0003] At present, there are mainly two methods for monitoring and detecting heavy metal pollutants: one is physical and chemical analysis methods, such as inductively coupled plasma atomic emission spectroscopy (ICP-AES), inductively coupled plasma mass spectrometry (ICP-MS), etc., its advantages are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12Q1/66C12Q1/02G01N21/64C12R1/19
Inventor 庄国强侯启会马安周
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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