Construction of soil environmental DNA cosmid libraries and screening for clones that produce biologically active small molecules.
Culture-independent studies on environmental samples indicate that most bacteria are not readily cultured in the laboratory. The small fraction of bacteria that have been successfully cultured from environmental samples have been a very rewarding source of novel biologically active natural products. The introduction of DNA extracted directly from environmental samples into easily cultured bacteria and the screening of these large libraries for clones that produce biologically active small molecules is one means to access natural products encoded by the genomes of previously uncultured bacteria. This protocol provides detailed procedures for cloning DNA directly from environmental samples and screening these clones for the production of antibacterially active natural products. The entire protocol, from soil sample to the identification of antibacterially active environmental DNA clones, will take approximately 2 weeks.
Brady
Sean F
SF
Laboratory of Genetically Encoded Small Molecules, The Rockefeller University, 1230 York Avenue, New York, New York 10021, USA. sbrady@rockefeller.edu
D000900:Anti-Bacterial Agents / Q000302:isolation & purification*; D001419:Bacteria / Q000235:genetics*; D001688:Biological Products / Q000302:isolation & purification*; D003001:Cloning, Molecular / Q000379:methods; D003360:Cosmids / Q000235:genetics*; D015723:Gene Library; D012988:Soil Microbiology
D000900:Anti-Bacterial Agents; D001688:Biological Products