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Construction of a sensitive and specific lead biosensor using a genetically engineered bacterial system with a luciferase gene reporter controlled by pbr and cadA promoters
Reza Nedaeinia
Isfahan University of Medical Sciences
Corresponding Author
ORCiD: https://orcid.org/0000-0001-9922-7181
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at BioMedical Engineering OnLine.
Background: A bacterial biosensor refers to genetically engineered bacteria that produce an assessable signal in the presence of a physical or chemical agent in the environment.
Methods: We have designed and evaluated a bacterial biosensor expressing a luciferase-reporter gene controlled by pbr and cadA promoters in Cupriavidus metallidurans (previously termed Ralstonia metallidurans) containing the CH34 and pI258 plasmids of Staphylococcus aureus, respectively, and that can be used for the detection of heavy metals. In the present study, we have produced and evaluated biosensor plasmids designated pGL3-luc/pbr-biosensor and pGL3-luc/cad-biosensor, that were based on the expression of luc+ under the control of the cad promoter and the cadC gene of S. aureus plasmid pI258 and pbr promoter and pbrR gene from plasmid pMOL30 of Cupriavidus metallidurans.
Results: We found that the pGL3-luc/pbr-biosensor may be used to measure lead concentrations between 1-100 μM in the presence of other metals, including: zinc, cadmium, tin and nickel. The latter metals did not result in any significant signal. The pGL3-luc/cad-biosensor could detect lead concentrations between 10 nM to 10 μM.
Conclusions: This biosensor was found to be a specific for measuring lead ions in both environmental and biological samples.
Keywords
Lead detection, Bacterial biosensor, Pollution control system
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CURRENT STATUS: Published
Published
On 19 Oct, 2020
No community comments so far
Editorial decision: Accept
On 05 Sep, 2020
Editor assigned
On 02 Sep, 2020
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On 01 Sep, 2020
Editor invited
On 01 Sep, 2020
No comments provided
Editorial decision: Minor revision
On 30 Aug, 2020
Editor assigned
On 29 Aug, 2020
Submission checks complete
On 28 Aug, 2020
Editor invited
On 28 Aug, 2020
No comments provided
Editorial decision: Minor revision
On 01 Aug, 2020
Editor assigned
On 29 Jul, 2020
Submission checks complete
On 28 Jul, 2020
Editor invited
On 28 Jul, 2020
Version 4
Posted 22 Jun, 2020
No comments provided
Editorial decision: Major revision
On 21 Jun, 2020
Editor assigned
On 19 Jun, 2020
Submission checks complete
On 18 Jun, 2020
Editor invited
On 18 Jun, 2020
No comments provided
Editor assigned
On 18 May, 2020
Editorial decision: Minor revision
On 18 May, 2020
Submission checks complete
On 17 May, 2020
Editor invited
On 17 May, 2020
No comments provided
Editorial decision: Major revision
On 18 Apr, 2020
Reviewer #4 agreed
On 10 Apr, 2020
Review #2 received
Received 09 Apr, 2020
Reviewer #3 agreed
On 07 Apr, 2020
Review #1 received
Received 31 Mar, 2020
Reviewer #2 agreed
On 30 Mar, 2020
Reviewer #1 agreed
On 27 Mar, 2020
Reviewers invited
Invitations sent on 27 Mar, 2020
Editor assigned
On 26 Mar, 2020
Submission checks complete
On 25 Mar, 2020
Editor invited
On 25 Mar, 2020
No comments provided
Editorial decision: Revise before peer review
On 19 Mar, 2020
Editor assigned
On 18 Mar, 2020
First submitted
On 17 Mar, 2020
Submission checks complete
On 17 Mar, 2020
Editor invited
On 17 Mar, 2020
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This preprint is under consideration at BioMedical Engineering OnLine. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Construction of a sensitive and specific lead biosensor using a genetically engineered bacterial system with a luciferase gene reporter controlled by pbr and cadA promoters
Reza Nedaeinia
Isfahan University of Medical Sciences
Corresponding Author
ORCiD: https://orcid.org/0000-0001-9922-7181
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
Published
On 19 Oct, 2020
No community comments so far
Editorial decision: Accept
On 05 Sep, 2020
Editor assigned
On 02 Sep, 2020
Submission checks complete
On 01 Sep, 2020
Editor invited
On 01 Sep, 2020
No comments provided
Editorial decision: Minor revision
On 30 Aug, 2020
Editor assigned
On 29 Aug, 2020
Submission checks complete
On 28 Aug, 2020
Editor invited
On 28 Aug, 2020
No comments provided
Editorial decision: Minor revision
On 01 Aug, 2020
Editor assigned
On 29 Jul, 2020
Submission checks complete
On 28 Jul, 2020
Editor invited
On 28 Jul, 2020
Version 4
Posted 22 Jun, 2020
No comments provided
Editorial decision: Major revision
On 21 Jun, 2020
Editor assigned
On 19 Jun, 2020
Submission checks complete
On 18 Jun, 2020
Editor invited
On 18 Jun, 2020
No comments provided
Editor assigned
On 18 May, 2020
Editorial decision: Minor revision
On 18 May, 2020
Submission checks complete
On 17 May, 2020
Editor invited
On 17 May, 2020
No comments provided
Editorial decision: Major revision
On 18 Apr, 2020
Reviewer #4 agreed
On 10 Apr, 2020
Review #2 received
Received 09 Apr, 2020
Reviewer #3 agreed
On 07 Apr, 2020
Review #1 received
Received 31 Mar, 2020
Reviewer #2 agreed
On 30 Mar, 2020
Reviewer #1 agreed
On 27 Mar, 2020
Reviewers invited
Invitations sent on 27 Mar, 2020
Editor assigned
On 26 Mar, 2020
Submission checks complete
On 25 Mar, 2020
Editor invited
On 25 Mar, 2020
No comments provided
Editorial decision: Revise before peer review
On 19 Mar, 2020
Editor assigned
On 18 Mar, 2020
First submitted
On 17 Mar, 2020
Submission checks complete
On 17 Mar, 2020
Editor invited
On 17 Mar, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Biochemical Research Methods
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at BioMedical Engineering OnLine.
Background: A bacterial biosensor refers to genetically engineered bacteria that produce an assessable signal in the presence of a physical or chemical agent in the environment.
Methods: We have designed and evaluated a bacterial biosensor expressing a luciferase-reporter gene controlled by pbr and cadA promoters in Cupriavidus metallidurans (previously termed Ralstonia metallidurans) containing the CH34 and pI258 plasmids of Staphylococcus aureus, respectively, and that can be used for the detection of heavy metals. In the present study, we have produced and evaluated biosensor plasmids designated pGL3-luc/pbr-biosensor and pGL3-luc/cad-biosensor, that were based on the expression of luc+ under the control of the cad promoter and the cadC gene of S. aureus plasmid pI258 and pbr promoter and pbrR gene from plasmid pMOL30 of Cupriavidus metallidurans.
Results: We found that the pGL3-luc/pbr-biosensor may be used to measure lead concentrations between 1-100 μM in the presence of other metals, including: zinc, cadmium, tin and nickel. The latter metals did not result in any significant signal. The pGL3-luc/cad-biosensor could detect lead concentrations between 10 nM to 10 μM.
Conclusions: This biosensor was found to be a specific for measuring lead ions in both environmental and biological samples.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7