Construction of mercury ion microorganism whole-cell biosensor taking violacein as output signal and application thereof

A violacein and mercury ion technology, applied in the biological field, can solve the problems that have not yet been reported by biosensors

Pending Publication Date: 2021-08-10
SHENZHEN PREVENTION & TREATMENT CENT FOR OCCUPATIONAL DISEASES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, there is no relevant report on the mercury ion microbial whole-cell biosensor with violacein as the output signal

Method used

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  • Construction of mercury ion microorganism whole-cell biosensor taking violacein as output signal and application thereof
  • Construction of mercury ion microorganism whole-cell biosensor taking violacein as output signal and application thereof
  • Construction of mercury ion microorganism whole-cell biosensor taking violacein as output signal and application thereof

Examples

Experimental program
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Embodiment 1

[0067] Example 1. Construction and application of mercury ion microbial whole-cell biosensor with violacein as output signal

[0068] 1. Construction of recombinant plasmid pPmer-vio

[0069] The first step is to use pET-21a(+) as the vector to synthesize the violacein synthetic gene module (position 555-7924 ​​of SEQ ID No.1), and insert it into pET-21a(+) using NdeI / SacI In , the recombinant plasmid verified by sequencing was named pET-vio.

[0070] The structure of pET-vio is described as: a recombinant plasmid in which the small fragment between NdeI / SacI of the pET-21a(+) vector is replaced with the DNA fragment shown in the 555-7924th position of SEQID No.1.

[0071] In the second step, the merR-mer promoter gene module (position 7-512 of SEQ ID No.1) is synthesized from the entire gene, inserted into the pET-vio vector using BglII / XbaI, and the recombinant plasmid verified by sequencing is named as pPmer-vio.

[0072] The structure of pPmer-vio is described as: a rec...

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Abstract

The invention discloses construction of a mercury ion microorganism whole-cell biosensor taking violacein as an output signal and an application thereof. The invention provides a recombinant bacterium which is obtained by introducing a recombinant vector A into a recipient bacterium, the recombinant vector A contains a DNA fragment A; the DNA fragment A contains a mer bidirectional promoter, one side of the mer bidirectional promoter is an MerR protein coding gene, and the other side of the mer bidirectional promoter is a violacein synthesis gene module. The recombinant bacterium provided by the invention can be used as a mercury ion microbial whole-cell biosensor taking violacein as an output signal. On one hand, qualitative indication of exposure of the target heavy metal can be achieved through change of cell colors, and on the other hand, intracellular violacein can be extracted through an organic solvent, and quantitative analysis of the target heavy metal is conducted through colorimetry.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the construction and application of a mercury ion microbial whole-cell biosensor with violacein as an output signal. Background technique [0002] Mercury is an environmental pollutant that exists in the atmosphere for a long time and has global mobility. Human beings have been aware of the toxicity of mercury for several centuries. In the 1950s, an event that had a significant impact on human health due to environmental exposure to mercury occurred, that is, the Minamata disease event in Japan. Mercury exists in several different chemical forms, including elemental mercury, organic mercury, and inorganic mercury. Among them, Hg(II) can be directly ingested by biological cells and metabolized into methylmercury, the most toxic organic form. Whole-cell biosensors directly simulate environmental microorganisms to detect bioavailable target metals, making them irreplaceable in environ...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/70C12Q1/04G01N21/78C12R1/19
CPCC12N15/70C07K14/245C12Q1/04G01N21/78
Inventor 惠长野郭妍杨学琴林怡然黄红英李丽梅
Owner SHENZHEN PREVENTION & TREATMENT CENT FOR OCCUPATIONAL DISEASES
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