Escherichia coli ribose binding protein based bioreporters revisited
| Autor: | Artur Reimer, Shantanu Roy, Sharon Yagur-Kroll, Jan Roelof van der Meer, Shimshon Belkin |
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| Rok vydání: | 2014 |
| Předmět: |
0303 health sciences
Multidisciplinary 030306 microbiology Binding protein Biology musculoskeletal system medicine.disease_cause biology.organism_classification In vitro 03 medical and health sciences chemistry.chemical_compound chemistry Biochemistry Escherichia Periplasmic Binding Proteins Ribose medicine Bioreporter Escherichia coli Bacteria 030304 developmental biology |
| Zdroj: | Scientific Reports, vol. 4, pp. 5626 |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/srep05626 |
| Popis: | Bioreporter bacteria, i.e., strains engineered to respond to chemical exposure by production of reporter proteins, have attracted wide interest because of their potential to offer cheap and simple alternative analytics for specified compounds or conditions. Bioreporter construction has mostly exploited the natural variation of sensory proteins, but it has been proposed that computational design of new substrate binding properties could lead to completely novel detection specificities at very low affinities. Here we reconstruct a bioreporter system based on the native Escherichia coli ribose binding protein RbsB and one of its computationally designed variants, reported to be capable of binding 2,4,6-trinitrotoluene (TNT). Our results show in vivo reporter induction at 50 nM ribose, and a 125 nM affinity constant for in vitro ribose binding to RbsB. In contrast, the purified published TNT-binding variant did not bind TNT nor did TNT cause induction of the E. coli reporter system. |
| Databáze: | OpenAIRE |
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