Open AccessFluorescence Resonance Energy Transfer (FRET) based molecular detection of a genetically modified PCB degrader in soil
Author(s) -
Hogan Jill,
Sherlock Orla,
Ryan David,
Whelan Clare,
Francesconi Stephen,
Rivilla Rafael,
Dowling David N
Publication year - 2004
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2004.tb09668.x
Subject(s) - förster resonance energy transfer , pseudomonas fluorescens , rhizosphere , bacteria , sybr green i , fluorescence , biology , detection limit , operon , recombinant dna , dna , microbiology and biotechnology , chemistry , genetics , real time polymerase chain reaction , mutant , chromatography , gene , physics , quantum mechanics
Genetic analysis of the location of a mini‐Tn5 promoted insertion of the LB400 bph operon in the rhizosphere coloniser Pseudomonas fluorescens F113rifPCB, allowed the development of a specific PCR detection system based on the unique DNA sequence at this insertion site. Real time PCR using both SYBR green chemistry and Fluorescence Resonance Energy Transfer probes allowed the precise identification of the recombinant strain and its quantitative detection in soil microcosms over a (bacteria/g) range of five orders of magnitude. This new assay can detect the genetically modified microorganism from soil in less than 90 min and at levels below the detection limits of standard PCR or cultivable counts on selective media.