Detection of single-copy functional genes in prokaryotic cells by two-pass TSA-FISH with polynucleotide probes

Access this Article

Search this Article

Abstract

In situ detection of functional genes with single-cell resolution is currently of interest to microbiologists. Here, we developed a two-pass tyramide signal amplification (TSA)-fluorescence in situ hybridization (FISH) protocol with PCR-derived polynucleotide probes for the detection of single-copy genes in prokaryotic cells. The mcrA gene and the apsA gene in methanogens and sulfate-reducing bacteria, respectively, were targeted. The protocol showed bright fluorescence with a good signal-to-noise ratio and achieved a high efficiency of detection (>98%). The discrimination threshold was approximately 82-89% sequence identity. Microorganisms possessing the mcrA or apsA gene in anaerobic sludge samples were successfully detected by two-pass TSA-FISH with polynucleotide probes. The developed protocol is useful for identifying single microbial cells based on functional gene sequences.

Journal

  • Journal of Microbiological Methods

    Journal of Microbiological Methods 88(2), 218-223, 2012

    Elsevier Science BV

Cited by:  1

Codes

  • NII Article ID (NAID)
    120005308241
  • NII NACSIS-CAT ID (NCID)
    AA1063956X
  • Text Lang
    ENG
  • Article Type
    Journal Article
  • ISSN
    0167-7012
  • Data Source
    CJPref  IR 
Page Top