Novel genes retrieved from environmental DNA by polymerase chain reaction: current genome-walking techniques for future metagenome applications.
Environmental DNA is an extremely rich source of genes encoding enzymes with novel biocatalytic activities. To tap this source, function-based and sequence-based strategies have been established to isolate, clone, and express these novel metagenome-derived genes. Sequence-based strategies, which rely on PCR with consensus primers and genome walking, represent an efficient and inexpensive alternative to activity-based screening of recombinant strains harbouring fragments of environmental DNA. This review covers the diverse array of genome-walking techniques, which were originally developed for genomic DNA and currently are also used for PCR-based recovery of entire genes from the metagenome. These sequence-based gene mining methods appear to offer a powerful tool for retrieving from the metagenome novel genes encoding biocatalysts with potential applications in biotechnology.
Kotik
Michael
M
Laboratory of Enzyme Technology, Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague 4, Czech Republic. kotik@biomed.cas.cz
D016386:Chromosome Walking / Q000379:methods*; D004247:DNA / Q000235:genetics*; D004777:Environment; D005796:Genes / Q000235:genetics*; D054892:Metagenome / Q000235:genetics*; D056186:Metagenomics / Q000379:methods*; D016133:Polymerase Chain Reaction
D004247:DNA