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Development of a PCR for the detection and identification of cyanobacterial nifD genes.
J Microbiol Methods. 2007 Sep; 70(3):550-6.JM

Abstract

In this study we have designed degenerate primers after comparative analysis of nifD gene sequences from public databases, and developed a PCR protocol for the amplification of nifD sequences from cyanobacteria. The primers were tested on a variety of nitrogenase-containing and nitrogenase-lacking bacteria. By using this protocol, we amplified nifD sequences from DNA that was isolated from three phototrophic microbial communities. Denaturing gradient gel electrophoresis (DGGE) and clone library analysis of the nifD amplicons showed the presence of distinct groups of diazotrophic cyanobacteria in each of the investigated microbial communities. Phylogenetic trees constructed from the sequences of nifD gene fragments are congruent with those based on ribosomal RNA gene sequences.

Authors+Show Affiliations

Department of Biotechnology, Delft University of Technology, NL-2628 BC Delft, The Netherlands.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17681622

Citation

Roeselers, Guus, et al. "Development of a PCR for the Detection and Identification of Cyanobacterial nifD Genes." Journal of Microbiological Methods, vol. 70, no. 3, 2007, pp. 550-6.
Roeselers G, Stal LJ, van Loosdrecht MC, et al. Development of a PCR for the detection and identification of cyanobacterial nifD genes. J Microbiol Methods. 2007;70(3):550-6.
Roeselers, G., Stal, L. J., van Loosdrecht, M. C., & Muyzer, G. (2007). Development of a PCR for the detection and identification of cyanobacterial nifD genes. Journal of Microbiological Methods, 70(3), 550-6.
Roeselers G, et al. Development of a PCR for the Detection and Identification of Cyanobacterial nifD Genes. J Microbiol Methods. 2007;70(3):550-6. PubMed PMID: 17681622.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of a PCR for the detection and identification of cyanobacterial nifD genes. AU - Roeselers,Guus, AU - Stal,Lucas J, AU - van Loosdrecht,Mark C M, AU - Muyzer,Gerard, Y1 - 2007/06/30/ PY - 2007/04/19/received PY - 2007/06/08/revised PY - 2007/06/20/accepted PY - 2007/8/8/pubmed PY - 2007/12/14/medline PY - 2007/8/8/entrez SP - 550 EP - 6 JF - Journal of microbiological methods JO - J Microbiol Methods VL - 70 IS - 3 N2 - In this study we have designed degenerate primers after comparative analysis of nifD gene sequences from public databases, and developed a PCR protocol for the amplification of nifD sequences from cyanobacteria. The primers were tested on a variety of nitrogenase-containing and nitrogenase-lacking bacteria. By using this protocol, we amplified nifD sequences from DNA that was isolated from three phototrophic microbial communities. Denaturing gradient gel electrophoresis (DGGE) and clone library analysis of the nifD amplicons showed the presence of distinct groups of diazotrophic cyanobacteria in each of the investigated microbial communities. Phylogenetic trees constructed from the sequences of nifD gene fragments are congruent with those based on ribosomal RNA gene sequences. SN - 0167-7012 UR - https://www.unboundmedicine.com/medline/citation/17681622/Development_of_a_PCR_for_the_detection_and_identification_of_cyanobacterial_nifD_genes_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0167-7012(07)00222-9 DB - PRIME DP - Unbound Medicine ER -