Abstract
We have developed techniques to clone the entire microbial metagenome; viruses, prokaryotes and eukaryotes. Established approaches were used to isolate environmental DNA and make prokaryotic gene libraries. These libraries contain genes encoding enzymes; cellulases and esterases have been functionally expressed. PCR amplification of environmental DNA has been used to identify integron associated gene cassettes encoding unidentified ORFs. Sequence independent DNA amplification was used to make a faecal virus metagenomic library. Analysis of this library showed the presence of protein ORFS especially enzymes involved in nucleic acid metabolism. Most virus ORFs were unrelated to known sequences. Finally metagenomic microbial eukaryotic RNA was reverse transcribed to select for mRNA; the cDNA was used to make libraries containing many eukaryotic ORFs.
