A metal-repressed promoter from gram-positive Bacillus subtilis is highly active and metal-induced in gram-negative Cupriavidus metallidurans
| Autor: | Ronaldo Biondo, Ana Clara Guerrini Schenberg, Elisabete José Vicente, Gabriela Ribeiro-dos-Santos, Oeber de Freitas Quadros |
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| Rok vydání: | 2010 |
| Předmět: |
Transcriptional Activation
Green Fluorescent Proteins lac operon Bioengineering Bacillus subtilis medicine.disease_cause Applied Microbiology and Biotechnology Fluorescence Microbiology Bacterial Proteins Genes Reporter Gene expression medicine Gene Silencing Promoter Regions Genetic Escherichia coli Reporter gene Bacillaceae biology Cupriavidus metallidurans Cupriavidus MICROBIOLOGIA Flow Cytometry biology.organism_classification Molecular biology Enterobacteriaceae DNA-Binding Proteins Lac Operon Metals Biotechnology |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| Popis: | A synthetic version of the metal-regulated gene A (mrgA) promoter from Bacillus subtilis, which in this Gram-positive bacterium is negatively regulated by manganese, iron, cobalt, or copper turned out to promote high level of basal gene expression that is further enhanced by Co(II), Cd(II), Mn(II), Zn(II), Cu(II), or Ni(II), when cloned in the Gram-negative bacterium Cupriavidus metallidurans. Promoter activity was monitored by expression of the reporter gene coding for the enhanced green fluorescent protein (EGFP), and cellular intensity fluorescence was quantified by flow cytometry. Expression levels in C. metallidurans driven by the heterologous promoter, here called pan, ranged from 20- to 53-fold the expression level driven by the Escherichia coli lac promoter (which is constitutively expressed in C. metallidurans), whether in the absence or presence of metal ions, respectively. The pan promoter did also function in E. coli in a constitutive pattern, regardless of the presence of Mn(II) or Fe(II). In conclusion, the pan promoter proved to be a powerful tool to express heterologous proteins in Gram-negative bacteria, especially in C. metallidurans grown upon high levels of toxic metals, with potential applications in bioremediation. |
| Databáze: | OpenAIRE |
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