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AssigneeTerragen Diversity, Inc.
InventorsBarbara Waters, Vivian Miao, Yap Wai Ho, Seow Kah Tong
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Degenerate primers which hybridize with various classes of antibiotic biosynthesis genes were used to amplify fragments of DNA from soil and lichen extracts. Cloning and sequencing of the amplified products showed that these products included a variety of novel and previously uncharacterized antibiotic biosynthesis gene sequences, the products of which have the potential to be active as antibiotics, immunosuppressors, antitumor agents, etc. Thus, antibiotic biosynthesis genes can be recovered from soil or lichens by a combining a sample with a pair of amplification primers under conditions suitable for polymerase chain reaction amplification, wherein the primer set is a degenerate primer set selected to hybridize with conserved regions of known antibiotic biosynthetic pathway genes, for example Type I and Type II polyketide synthase genes, isopenicillin N synthase genes, and peptide synthetase genes; cycling the combined sample through a plurality of amplification cycles to amplify DNA complementary to the primer set; and isolating the amplified DNA.
A method for recovery of DNA comprising a nucleotide sequence of a portion of a gene that encodes a protein that is involved in antibiotic synthesis, from humic materials or lichen, comprising the steps of:
An oligonucleotide primer consisting essentially of the sequence as defined in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7 or SEQ ID NO:8.
A composition comprising two oligonucleotide primers, said oligonucleotide primers each consisting essentially of the sequence as defined in (i) SEQ ID NO:
Dependencies are inferred from the claim text.