Use of degenerate oligonucleotides for amplification of the nifH gene from the marine cyanobacterium Trichodesmium thiebautii.

Applied and environmental microbiology ·Vol. 55 ·No. 10 ·1989-10-00 ·Pages 2522-6

Zehr JP, McReynolds LA

Abstract

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.

MeSH Terms

Amino Acid Sequence Base Sequence Cloning, Molecular Cyanobacteria/genetics DNA/genetics Gene Amplification Genes, Bacterial Molecular Sequence Data Nitrogen Fixation/genetics Oligodeoxyribonucleotides Polymerase Chain Reaction Sequence Homology, Nucleic Acid

Chemicals

Oligodeoxyribonucleotides DNA

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Zehr J P

New England BioLabs, Inc., Beverly, Massachusetts 01915.

McReynolds L A

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23 references, click to expand

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Article Info

Journal

Applied and environmental microbiology

Abbr.

Appl Environ Microbiol

ISSN

0099-2240

DOI

10.1128/aem.55.10.2522-2526.1989

Published

1989-10-00

Pages

2522-6

Language

English

Region

United States

NLM ID

7605801

PMCID

PMC203115

Subset

Databases

GENBANK

M29709

External Links

PubMed source DOI: 10.1128/aem.55.10.2522-2526.1989 PMC Full Text View journal details

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Use of degenerate oligonucleotides for amplification of the nifH gene from the marine cyanobacterium Trichodesmium thiebautii.

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