Journal article
Detection of an alkene monooxygenase in vinyl chloride-oxidizing bacteria with GeneFISH
Journal of microbiological methods, Vol.181, pp.106147-106147
02/2021
DOI: 10.1016/j.mimet.2021.106147
PMID: 33493490
Abstract
Fluorescence in situ hybridization (FISH) can provide information on the morphology, spatial arrangement, and local environment of individual cells enabling the investigation of intact microbial communities. GeneFISH uses polynucleotide probes and enzymatic signal amplification to detect genes that are present in low copy numbers. Previously, this technique has only been applied in a small number of closely related organisms. However, many important functional genes, such as those involved in xenobiotic degradation or pathogenesis, are present in diverse microbial strains. Here, we present a geneFISH method for the detection of the functional gene etnC, which encodes the alpha subunit of an alkene monooxygenase used by aerobic ethene and vinyl chloride oxidizing bacteria (etheneotrophs). The probe concentration was optimized and found to be 100 pg/μl, similar to previous geneFISH reports. Permeabilization was necessary for successful geneFISH labeling of Mycobacteria; sequential treatment with lysozyme and achromopeptidase was the most effective treatment. This method was able to detect etnC in several organisms including Mycobacteria and Nocardioides, demonstrating for the first time that a single geneFISH probe can detect a variety of alleles (>80% sequence similarity) across multiple species. Detection of etnC with geneFISH has practical applications for bioremediation. This method can be readily adapted for other functional genes and has broad applications for investigating microbial communities in natural and engineered systems.
Details
- Title: Subtitle
- Detection of an alkene monooxygenase in vinyl chloride-oxidizing bacteria with GeneFISH
- Creators
- Patrick M Richards - Department of Civil and Environmental Engineering, 4105 Seamans Center, The University of Iowa, Iowa City, IA 52242, USATimothy E Mattes - Department of Civil and Environmental Engineering, 4105 Seamans Center, The University of Iowa, Iowa City, IA 52242, USA. Electronic address: tim-mattes@uiowa.edu
- Resource Type
- Journal article
- Publication Details
- Journal of microbiological methods, Vol.181, pp.106147-106147
- Publisher
- Netherlands
- DOI
- 10.1016/j.mimet.2021.106147
- PMID
- 33493490
- ISSN
- 0167-7012
- eISSN
- 1872-8359
- Grant note
- DOI: 10.13039/100000146, name: Division of Chemical, Bioengineering, Environmental, and Transport Systems
- Language
- English
- Date published
- 02/2021
- Academic Unit
- Civil and Environmental Engineering
- Record Identifier
- 9984077373302771
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