A rapid reporter system using GFP as a reporter protein for identification and screening of synthetic stationary-phase promoters in Escherichia coli.
Appl Microbiol Biotechnol
; 70(2): 229-36, 2006 Mar.
Article
in En
| MEDLINE
| ID: mdl-16012833
ABSTRACT
To develop a rapid reporter system for the screening of stationary-phase promoters in Escherichia coli, the expression pattern of the green fluorescent protein (GFP) during bacterial cultivation was compared with that of the commonly used beta-galactosidase. Using GFP with enhanced fluorescence, the expression pattern of both reporter systems GFP and beta-galactosidase were similar and showed a typical induction of gene activity of the reporter genes, i.e. increase of expression at the transition from exponential to stationary phase. The expression was affected by the culture medium, i.e. in contrast to the complex medium (LB medium), the stationary-phase specific induction was only observed in synthetic medium (M9) when amino acids were added, whereas there was generally no induction in MOPS medium. To develop a rapid screening method on agar plates for stationary-phase promoters, a photographic approach was used, continued with computational image treatment. A screening method is presented which enables an on-line monitoring of gene activity.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Promoter Regions, Genetic
/
Genes, Reporter
/
Green Fluorescent Proteins
/
Escherichia coli
Type of study:
Diagnostic_studies
/
Evaluation_studies
/
Prognostic_studies
/
Screening_studies
Language:
En
Journal:
Appl Microbiol Biotechnol
Year:
2006
Document type:
Article
Affiliation country:
Alemania