Detection of aromatic catabolic gene expression in heterogeneous organic matter used for reduction of volatile organic compounds (VOC) by biofiltration.
A qualitative procedure of purified DNA/RNA co-extraction from complex organic matter, used as biofilter support for removing volatile organic compounds, was set up and applied to detect xylene monooxygenase gene expression by RT-PCR. A DNA/RNA extraction protocol based on a combination of sample lyophilization pre-treatment and CTAB--phenol/chloroform extraction procedure was optimized for the recovery of purified nucleic acids [100-500 ng DNA (10 kb) and 0.5-2 microg of rRNA 16S from 100 mg matrix]. PCR and RT-PCR protocols were established to detect xylene monooxygenase gene expression starting from differentially induced organic matrices obtained by biofiltration technology. This work allowed the microbial degradation activities in heterogeneous organic solid media to be studied and suggests a rapid method to follow specific biological activities during solid and/or semisolid organic substrates biotransformation.
Tell
Gianluca
G
Department of Biomedical Sciences & Technologies, Molecular Biology Section, University of Udine, Udine, Italy.
Paron
Igor
I
Civilini
Marcello
M
D000391:Air Microbiology; D001673:Biodegradation, Environmental; D004269:DNA, Bacterial / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods*; D006841:Hydrocarbons, Aromatic / Q000378:metabolism*; D008660:Metabolism / Q000502:physiology; D009930:Organic Chemicals / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D004269:DNA, Bacterial; D006841:Hydrocarbons, Aromatic; D009930:Organic Chemicals