Metaproteomic and metagenomic analyses of defined oceanic microbial populations using microwave cell fixation and flow cytometric sorting.
International audience A major obstacle in the molecular investigation of natural, especially oceanic, microbial cells is their adequate preservation for further land-based molecular analyses. Here, we examined the use of microwaves for cell fixation before high-speed flow cytometric sorting to defi...
Published in: | FEMS Microbiology Ecology |
---|---|
Main Authors: | , , , , , , , , , , |
Other Authors: | , , , , , , , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
HAL CCSD
2010
|
Subjects: | |
Online Access: | https://hal.archives-ouvertes.fr/hal-00528837 https://doi.org/10.1111/j.1574-6941.2010.00927.x |
id |
ftccsdartic:oai:HAL:hal-00528837v1 |
---|---|
record_format |
openpolar |
spelling |
ftccsdartic:oai:HAL:hal-00528837v1 2023-05-15T17:33:11+02:00 Metaproteomic and metagenomic analyses of defined oceanic microbial populations using microwave cell fixation and flow cytometric sorting. Mary, Isabelle Oliver, Anna Skipp, Paul Holland, Ross Topping, Juliette Tarran, Glen Scanlan, David J O'Connor, C David Whiteley, Andrew S Burkill, Peter H Zubkov, Mikhail V National Oceanography Centre (NOC) Laboratoire Microorganismes : Génome et Environnement (LMGE) Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Université d'Auvergne - Clermont-Ferrand I (UdA)-Centre National de la Recherche Scientifique (CNRS) Molecular Microbial Ecology Section Biodiversity and Ecosystem Function Group-CEH-Oxford School of Biological Sciences University of Southampton Plymouth Marine Laboratory Department of Biological Sciences Coventry University of Warwick Coventry 2010-10 https://hal.archives-ouvertes.fr/hal-00528837 https://doi.org/10.1111/j.1574-6941.2010.00927.x en eng HAL CCSD Wiley-Blackwell info:eu-repo/semantics/altIdentifier/doi/10.1111/j.1574-6941.2010.00927.x info:eu-repo/semantics/altIdentifier/pmid/20618860 hal-00528837 https://hal.archives-ouvertes.fr/hal-00528837 doi:10.1111/j.1574-6941.2010.00927.x PUBMED: 20618860 ISSN: 0168-6496 EISSN: 1574-6941 FEMS Microbiology Ecology https://hal.archives-ouvertes.fr/hal-00528837 FEMS Microbiology Ecology, Wiley-Blackwell, 2010, 74 (1), pp.10-8. ⟨10.1111/j.1574-6941.2010.00927.x⟩ [SDV.EE]Life Sciences [q-bio]/Ecology environment info:eu-repo/semantics/article Journal articles 2010 ftccsdartic https://doi.org/10.1111/j.1574-6941.2010.00927.x 2021-10-03T00:38:03Z International audience A major obstacle in the molecular investigation of natural, especially oceanic, microbial cells is their adequate preservation for further land-based molecular analyses. Here, we examined the use of microwaves for cell fixation before high-speed flow cytometric sorting to define the metaproteomes and metagenomes of key microbial populations. The microwave fixation procedure was established using cultures of Synechococcus cyanobacteria, the photosynthetic eukaryote Micromonas pusilla and the gammaproteobacterium Halomonas variabilis. Shotgun proteomic analyses showed that the profile of microwave-fixed and -unfixed Synechococcus sp. WH8102 cells was the same, and hence proteome identification of microwave-fixed sorted cells by nanoLC-MS/MS is possible. Microwave-fixed flow-sorted Synechococcus cells can also be successfully used for whole-genome amplification and fosmid library construction. We then carried out successful metaproteomic and metagenomic analyses of microwave-fixed Synechococcus cells flow sorted from concentrates of microbial cells, collected in the North Atlantic Ocean. Thus, the microwave fixation procedure developed appears to be useful for molecular studies of microbial populations in aquatic ecosystems. Article in Journal/Newspaper North Atlantic Archive ouverte HAL (Hyper Article en Ligne, CCSD - Centre pour la Communication Scientifique Directe) FEMS Microbiology Ecology 74 1 10 18 |
institution |
Open Polar |
collection |
Archive ouverte HAL (Hyper Article en Ligne, CCSD - Centre pour la Communication Scientifique Directe) |
op_collection_id |
ftccsdartic |
language |
English |
topic |
[SDV.EE]Life Sciences [q-bio]/Ecology environment |
spellingShingle |
[SDV.EE]Life Sciences [q-bio]/Ecology environment Mary, Isabelle Oliver, Anna Skipp, Paul Holland, Ross Topping, Juliette Tarran, Glen Scanlan, David J O'Connor, C David Whiteley, Andrew S Burkill, Peter H Zubkov, Mikhail V Metaproteomic and metagenomic analyses of defined oceanic microbial populations using microwave cell fixation and flow cytometric sorting. |
topic_facet |
[SDV.EE]Life Sciences [q-bio]/Ecology environment |
description |
International audience A major obstacle in the molecular investigation of natural, especially oceanic, microbial cells is their adequate preservation for further land-based molecular analyses. Here, we examined the use of microwaves for cell fixation before high-speed flow cytometric sorting to define the metaproteomes and metagenomes of key microbial populations. The microwave fixation procedure was established using cultures of Synechococcus cyanobacteria, the photosynthetic eukaryote Micromonas pusilla and the gammaproteobacterium Halomonas variabilis. Shotgun proteomic analyses showed that the profile of microwave-fixed and -unfixed Synechococcus sp. WH8102 cells was the same, and hence proteome identification of microwave-fixed sorted cells by nanoLC-MS/MS is possible. Microwave-fixed flow-sorted Synechococcus cells can also be successfully used for whole-genome amplification and fosmid library construction. We then carried out successful metaproteomic and metagenomic analyses of microwave-fixed Synechococcus cells flow sorted from concentrates of microbial cells, collected in the North Atlantic Ocean. Thus, the microwave fixation procedure developed appears to be useful for molecular studies of microbial populations in aquatic ecosystems. |
author2 |
National Oceanography Centre (NOC) Laboratoire Microorganismes : Génome et Environnement (LMGE) Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Université d'Auvergne - Clermont-Ferrand I (UdA)-Centre National de la Recherche Scientifique (CNRS) Molecular Microbial Ecology Section Biodiversity and Ecosystem Function Group-CEH-Oxford School of Biological Sciences University of Southampton Plymouth Marine Laboratory Department of Biological Sciences Coventry University of Warwick Coventry |
format |
Article in Journal/Newspaper |
author |
Mary, Isabelle Oliver, Anna Skipp, Paul Holland, Ross Topping, Juliette Tarran, Glen Scanlan, David J O'Connor, C David Whiteley, Andrew S Burkill, Peter H Zubkov, Mikhail V |
author_facet |
Mary, Isabelle Oliver, Anna Skipp, Paul Holland, Ross Topping, Juliette Tarran, Glen Scanlan, David J O'Connor, C David Whiteley, Andrew S Burkill, Peter H Zubkov, Mikhail V |
author_sort |
Mary, Isabelle |
title |
Metaproteomic and metagenomic analyses of defined oceanic microbial populations using microwave cell fixation and flow cytometric sorting. |
title_short |
Metaproteomic and metagenomic analyses of defined oceanic microbial populations using microwave cell fixation and flow cytometric sorting. |
title_full |
Metaproteomic and metagenomic analyses of defined oceanic microbial populations using microwave cell fixation and flow cytometric sorting. |
title_fullStr |
Metaproteomic and metagenomic analyses of defined oceanic microbial populations using microwave cell fixation and flow cytometric sorting. |
title_full_unstemmed |
Metaproteomic and metagenomic analyses of defined oceanic microbial populations using microwave cell fixation and flow cytometric sorting. |
title_sort |
metaproteomic and metagenomic analyses of defined oceanic microbial populations using microwave cell fixation and flow cytometric sorting. |
publisher |
HAL CCSD |
publishDate |
2010 |
url |
https://hal.archives-ouvertes.fr/hal-00528837 https://doi.org/10.1111/j.1574-6941.2010.00927.x |
genre |
North Atlantic |
genre_facet |
North Atlantic |
op_source |
ISSN: 0168-6496 EISSN: 1574-6941 FEMS Microbiology Ecology https://hal.archives-ouvertes.fr/hal-00528837 FEMS Microbiology Ecology, Wiley-Blackwell, 2010, 74 (1), pp.10-8. ⟨10.1111/j.1574-6941.2010.00927.x⟩ |
op_relation |
info:eu-repo/semantics/altIdentifier/doi/10.1111/j.1574-6941.2010.00927.x info:eu-repo/semantics/altIdentifier/pmid/20618860 hal-00528837 https://hal.archives-ouvertes.fr/hal-00528837 doi:10.1111/j.1574-6941.2010.00927.x PUBMED: 20618860 |
op_doi |
https://doi.org/10.1111/j.1574-6941.2010.00927.x |
container_title |
FEMS Microbiology Ecology |
container_volume |
74 |
container_issue |
1 |
container_start_page |
10 |
op_container_end_page |
18 |
_version_ |
1766131598635302912 |