Abstract
We have applied dual genetic selection to design a bacterial riboswitch with improved sensitivity by employing a high-affinity heterologous thiamine pyrophosphate (TPP) aptamer and modified selection conditions in Escherichia coli. The cells transformed with the engineered TPP riboswitches were characterized as whole-cell biosensors. The riboswitches were further studied in cell-free translation systems where they exhibited characteristics similar to those in vivo and a shorter response time for analysis. The flexibility of the riboswitch-based biosensors to accommodate different reporter genes was also demonstrated. Tuning of biosensor characteristics in vivo enables efficient development of aptamer-based whole-cell and cell-free biosensors.
Original language | English (US) |
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Pages (from-to) | 108-122 |
Number of pages | 15 |
Journal | Analytical Letters |
Volume | 42 |
Issue number | 1 |
DOIs | |
State | Published - Jan 2009 |
Keywords
- Cell-free biosensor
- Dual genetic selection
- Riboswitch
- Whole-cell biosensor
ASJC Scopus subject areas
- Analytical Chemistry
- Biochemistry
- Clinical Biochemistry
- Biochemistry, medical
- Electrochemistry
- Spectroscopy