Efficient Design Strategy for Whole-Cell and Cell-Free Biosensors based on Engineered Riboswitches

Autor:	Norihito Muranaka, Yohei Yokobayashi, Vandana Sharma, Yoko Nomura
Rok vydání:	2009
Předmět:	Riboswitch Reporter gene Chemistry Aptamer Biochemistry (medical) Clinical Biochemistry technology industry and agriculture Heterologous macromolecular substances Computational biology medicine.disease_cause Biochemistry Analytical Chemistry chemistry.chemical_compound In vivo Electrochemistry medicine Escherichia coli Biosensor Spectroscopy Thiamine pyrophosphate
Zdroj:	Analytical Letters. 42:108-122
ISSN:	1532-236X 0003-2719
DOI:	10.1080/00032710802568556
Popis:	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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::c8fb3de3ddbd53e486e44044a4e0d0e4 https://doi.org/10.1080/00032710802568556 Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
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