Construction and genetic improvement of copper bioreporter Escherichia Coli

Biomedical and Biotechnology Research Journal

关键词: Copper (D003300);    biosensors (D015374);    Luciferase (D008156);    Escherichia coli (D004926);    Vibrio fischeri (D048908);    Bioreporter;   

DOI  :  10.4103/bbrj.bbrj_65_17

来源: DOAJ

【 摘 要 】

Background: Copper is a pollutant compound which can cause earnest toxicity in human and some organisms. Bioreporters are frugal and non-toxic detectors for pollution compounds. Precedent designed recombinant Escherichia coli copper bioreporters with the lux gene of Vibrio fischeri or Aequorin luciferase of Jellyfish does not provide a high sensitivity. The aim of current study was to design an incipient Copper bioreporter with applying firefly luciferase and Copper resistance promoter of P. syringae pv.Tomato in Escherichia coli XL1-Blue. Methods: Recombinant pGL3 was obtained by applying the pGL3-Control vector to Escherichia coli XL1-Blue, by Polymerase Chain Reaction method and double digestion. Recombinant Escherichia coli cells were cultured with applying different concentrations of copper sulphate to study the activity of luciferase by Luminometer. Copper bioreporter specificity was resolute by different concentrations of Zinc sulfate and Ferric sulfate. Results: Recombinant Escherichia coli BL21, with copper promoter gene in pGL3 Vector showed the highest Luciferase activity in 0.1 millimolar of Copper sulfate. The highest Luciferase activity was in 0.09 millimolar and 1.0 millimolar of Zinc sulfate and Ferric sulfate respectively. Conclusion: Current study provided a categorical bioreporter for detecting copper, utilizing firefly luciferase with a high specificity (96.1%). By optimizing inhibitor factors, application of current copper bioreporter can be developed in human life.

【 授权许可】

Unknown