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

BioMedical Engineering OnLine

Maryam Ranjbar¹  Mozhdeh Nedaeinia²  Gordon A. Ferns³  Saman Hosseinkhani⁴  Mohammad Goli⁵  Mohammad Nourmohammadi⁶  Ali Mahmoudi⁷  Zahra Farjami⁷  Esmail Nourmohammadi⁸  Majid Sadeghizadeh⁹  Neshat Ebrahimi^1,10  Reza Nedaeinia^1,11  Hoda Khoshdel-Sarkarizi^1,12 

[1] Advanced Materials Research Center, Department of Materials Engineering, Najafabad Branch, Islamic Azad University, Najafabad, Iran;Applied Physiology Research Center, Isfahan University of Medical Sciences, Isfahan, Iran;Brighton and Sussex Medical School, Division of Medical Education, Falmer, BN1 9PH, Brighton, Sussex, UK;Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran;Department of Food Science and Technology, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran;Department of Health Research Center, Mashhad University of Medical Sciences, Mashhad, Iran;Department of Medical Biotechnology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran;Department of Medical Biotechnology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran;Research Center of Advanced Technologies in Medicine, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh, Iran;Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran;Laboratory of Cedars-Sinai Medical Center, Los Angeles, CA, USA;Pediatric Inherited Diseases Research Center, Research Institute for Primordial Prevention of Non-Communicable Disease, Isfahan University of Medical Sciences, Isfahan, Iran;Research Center of Advanced Technologies in Medicine, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh, Iran;

关键词: Lead;    Bacterial biosensor;    Pbr;    Luciferase;    cadA;   

DOI  :  10.1186/s12938-020-00816-w

来源: Springer

【 摘 要 】

BackgroundA bacterial biosensor refers to genetically engineered bacteria that produce an assessable signal in the presence of a physical or chemical agent in the environment.MethodsWe 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+ and 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.ResultsWe 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.ConclusionsThis biosensor was found to be specific for measuring lead ions in both environmental and biological samples.

【 授权许可】

CC BY