Use of degenerate oligonucleotides for amplification of the nifH gene from the marine cyanobacterium Trichodesmium thiebautii.
Trichodesmium spp. are marine filamentous, nonheterocystous, nitrogen-fixing cyanobacteria which are an important component of marine ecosystems. This organism has never been maintained in axenic culture, and there has remained some doubt as to the identity of the organism responsible for nitrogen fixation in Trichodesmium aggregates. By using degenerate oligonucleotide primers, it has been possible to amplify, clone, and sequence a segment of the nifH gene from a natural assemblage of Trichodesmium thiebautii. Examination of the DNA and presumed amino acid sequence shows that the gene is most closely related to that of Anabaena spp. and therefore is most likely a cyanobacterial nifH gene. The use of degenerate oligonucleotides, in concert with the polymerase chain reaction, can be a powerful tool for the cloning and sequencing of a variety of genes from microorganisms in the environment.
Zehr
J P
JP
New England BioLabs, Inc., Beverly, Massachusetts 01915.
McReynolds
L A
LA
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000235:genetics*; D004247:DNA / Q000235:genetics; D005784:Gene Amplification; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009586:Nitrogen Fixation / Q000235:genetics*; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid
D009838:Oligodeoxyribonucleotides; D004247:DNA