Metaproteomic and metagenomic analyses of defined oceanic microbial populations using microwave cell fixation and flow cytometric sorting
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 an...
Published in: | FEMS Microbiology Ecology |
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Online Access: | https://wrap.warwick.ac.uk/id/eprint/5240/ https://doi.org/10.1111/j.1574-6941.2010.00927.x |
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ftuwarwick:oai:wrap.warwick.ac.uk:5240 2024-09-30T14:39:35+00:00 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. 2010-10 https://wrap.warwick.ac.uk/id/eprint/5240/ https://doi.org/10.1111/j.1574-6941.2010.00927.x unknown Wiley-Blackwell Publishing Ltd. 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. and Zubkov, Mikhail V. (2010) Metaproteomic and metagenomic analyses of defined oceanic microbial populations using microwave cell fixation and flow cytometric sorting. FEMS Microbiology Ecology, Vol.74 (No.1). pp. 10-18. doi:10.1111/j.1574-6941.2010.00927.x <https://doi.org/10.1111/j.1574-6941.2010.00927.x> ISSN 0168-6496. 10.1111/j.1574-6941.2010.00927.x QR Microbiology Journal Article NonPeerReviewed 2010 ftuwarwick https://doi.org/10.1111/j.1574-6941.2010.00927.x 2024-09-05T05:23:56Z 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 The University of Warwick: WRAP - Warwick Research Archive Portal FEMS Microbiology Ecology 74 1 10 18 |
institution |
Open Polar |
collection |
The University of Warwick: WRAP - Warwick Research Archive Portal |
op_collection_id |
ftuwarwick |
language |
unknown |
topic |
QR Microbiology |
spellingShingle |
QR Microbiology 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 |
QR Microbiology |
description |
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. |
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 |
Wiley-Blackwell Publishing Ltd. |
publishDate |
2010 |
url |
https://wrap.warwick.ac.uk/id/eprint/5240/ https://doi.org/10.1111/j.1574-6941.2010.00927.x |
genre |
North Atlantic |
genre_facet |
North Atlantic |
op_relation |
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. and Zubkov, Mikhail V. (2010) Metaproteomic and metagenomic analyses of defined oceanic microbial populations using microwave cell fixation and flow cytometric sorting. FEMS Microbiology Ecology, Vol.74 (No.1). pp. 10-18. doi:10.1111/j.1574-6941.2010.00927.x <https://doi.org/10.1111/j.1574-6941.2010.00927.x> ISSN 0168-6496. 10.1111/j.1574-6941.2010.00927.x |
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_ |
1811642203734278144 |