Fluorescence-based cadmium ion concentration detection method by using whole cell biosensor

A biosensor, concentration detection technology, applied in biochemical equipment and methods, microbial determination/inspection, fluorescence/phosphorescence, etc., can solve the problems of complex and time-consuming sample pretreatment

Inactive Publication Date: 2014-03-12
PEKING UNIV SHENZHEN GRADUATE SCHOOL
View PDF0 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the above methods require expensive instruments and equipment, and the sample pretreatment is complex and time-consuming, the development of a low-cost, easy-to-operate, and rapid quantitative cadmium ion detection method is a work with practical application value.

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
  • Fluorescence-based cadmium ion concentration detection method by using whole cell biosensor
  • Fluorescence-based cadmium ion concentration detection method by using whole cell biosensor
  • Fluorescence-based cadmium ion concentration detection method by using whole cell biosensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0012] Preparation of Pseudomonas putida whole-cell biosensor:

[0013] The cadmium-specific operon sequence was amplified using polymerase chain reaction (PCR) technology (see nucleotide sequence listing). The operon sequence and the green fluorescent protein coding gene are constructed into a plasmid, so that the operon sequence directly regulates the expression of green fluorescent protein (see figure 1 ).

[0014] Transform the constructed plasmid into Pseudomonas putida as follows:

[0015] 1) Take the well-grown Pseudomonas putida bacteria liquid in the logarithmic phase, centrifuge at 3000×g for 5 minutes at 4°C, and discard the supernatant;

[0016] 2) Use 1 / 2 volume of pre-cooled 0.1M CaCl 2 Resuspend, centrifuge at 3000×g for 5 minutes at 4°C, discard the supernatant;

[0017] 3) Use 1 / 10 volume of 0.1M CaCl containing 20% ​​glycerol 2 Resuspend the bacteria to make Pseudomonas putida competent cells;

[0018] 4) Add 0.1-0.2 μg of the plasmid to 100 μl of com...

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 relates to a fluorescence-based cadmium ion concentration detection method by using whole cell biosensor, and provides whole cell biosensors used for detecting the concentration of cadmium ions, and a method for quantitatively detecting the concentration of cadmium ions in an aqueous solution by utilizing the biosensors. Whole cells are bacteria having a biological metabolism activity, and the bacteria used in the invention comprise Escherichia coli and Pseudomonas putida. The biosensors realizes the detection of the concentration of the cadmium ions through the steps of identifying cadmium ions by a transcription regulation protein CadR, combing with the cadmium ions, combining with a CadR combined operon sequence (SEQ2) containing the cadmium ions, starting fluorescin transcription, converting the amount of the cadmium ions to the fluorescin expression level, and detecting the fluorescin fluorescence intensity.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a technology for quantitatively detecting the concentration of cadmium ions in an aqueous solution by using a whole-cell sensor capable of synthesizing green fluorescent protein. Background technique [0002] Human industrial and agricultural production activities have caused cadmium to enter the environment in various forms, producing a large amount of cadmium-containing wastewater and causing serious pollution to the environment. For a long time, the detection and analysis methods of cadmium ion concentration in aqueous solution have mainly relied on chemical and physical means, including colorimetric analysis, atomic absorption spectrometry, electrochemical polarography, plasma emission spectrometry, etc. Since the above methods require expensive instruments and equipment, sample pretreatment is complicated and time-consuming, etc., the development of a low-cost, easy-to-op...

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): C12N1/21C12N15/63C12Q1/02G01N21/64
Inventor 陶虎春彭智文李鹏松雷涛魏雪艳苏捷于太安
Owner PEKING UNIV SHENZHEN GRADUATE SCHOOL
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