期刊
ANALYTICAL LETTERS
卷 42, 期 1, 页码 108-122出版社
TAYLOR & FRANCIS INC
DOI: 10.1080/00032710802568556
关键词
Cell-free biosensor; dual genetic selection; riboswitch; whole-cell biosensor
资金
- Direct For Computer & Info Scie & Enginr [0829536] Funding Source: National Science Foundation
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.
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