d-Xylose detection in Escherichia coli by a xylose binding protein-dependent response
| Autor: | Lucas Ferreira Ribeiro, Gilvan Pessoa Furtado, Richard J. Ward, Fabiana Fernandes Bressan, Flavio Meireles |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Arabinose
Green Fluorescent Proteins Population Bioengineering Xylose Biology medicine.disease_cause Applied Microbiology and Biotechnology Marker gene Green fluorescent protein chemistry.chemical_compound Escherichia coli medicine ESCHERICHIA Cloning Molecular Promoter Regions Genetic education Gene education.field_of_study Endo-1 4-beta Xylanases Proteins Gene Expression Regulation Bacterial General Medicine Molecular biology chemistry Biochemistry Carrier Proteins Xylose binding Bacillus subtilis Biotechnology |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| ISSN: | 0168-1656 |
| Popis: | A gene circuit for the controlled expression of a marker gene and for the assay of xylose concentration in Escherichia coli has been designed and tested. The xylF coding sequence for the xylose binding protein (XBP) was cloned in pT7T318U downstream from the promoter for xylanase A from B. subtilis (Pbsu), together with the GFP coding sequence (gfp) under the control of the xylF promoter, forming the pT7T3-GFP-XBP construct. GFP fluorescence in Escherichia coli JW3538-1 xylF-transformed with pT7T3-GFP-XBP was approximately 1.4 × higher after 520 min growth in the presence of 5mM xylose than in cells transformed with pT7T3-GFP. Under saturating xylose concentration, flow cytometry analysis showed that all cells resulted in homogeneous populations, and the population with XBP showed a fluorescence greater than that without XBP. Activity of the xylF promoter in cells transformed with pT7T3-GFP-XBP was ≈ 40% higher than with the pT7T3-GFP. No response was observed with arabinose and ribose, showing that the expression effects were specific for xylose, demonstrating the potential use of the gene circuit as a biosensor. |
| Databáze: | OpenAIRE |
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