Development of a highly specific and sensitive cadmium and lead microbial biosensor using synthetic CadC-T7 genetic circuitry

Biosens Bioelectron. 2016 May 15:79:701-8. doi: 10.1016/j.bios.2015.12.101. Epub 2015 Dec 29.

Abstract

Multiple copies of a cadC homolog encoding a heavy metal-responsive transcription factor were found in the genome of a bacterium isolated from ocean sediment, and the heavy metal responses of the encoded proteins were characterized using a fluorescence reporter assay. Each CadC regulator exhibited distinct specificity in response to heavy metal ions, indicating their potential use as modular heavy metal biosensors. Next, we constructed CadC-controlled T7 RNA transcription systems for intracellular signal amplification, i.e., higher sensitivity. Flow cytometry revealed that cadmium and lead ions could be recognized specifically by CadC-T7 biosensors, which could be combined with a microfluidic platform to generate heavy metal biosensor devices with increased sensitivity. Our results demonstrate the successful development of synthetic CadC-T7 genetic circuitry for use in improved heavy metal biosensor microfluidic devices.

Keywords: CadC; Heavy metal ions; Microbial biosensor; Microfluidic device; T7 RNA polymerase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacteria / chemistry
  • Bacteria / genetics*
  • Bacteria / metabolism
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Base Sequence
  • Biosensing Techniques / methods*
  • Cadmium / analysis*
  • Cadmium / metabolism
  • Gene Expression Regulation, Bacterial*
  • Lead / analysis*
  • Lead / metabolism
  • Microfluidic Analytical Techniques / methods
  • Molecular Sequence Data
  • Transcription Factors / chemistry
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • Bacterial Proteins
  • Transcription Factors
  • Cadmium
  • Lead