Abstract:
: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.
journal_name
FEMS Microbiol Ecoljournal_title
FEMS microbiology ecologyauthors
Mary I,Oliver A,Skipp P,Holland R,Topping J,Tarran G,Scanlan DJ,O'Connor CD,Whiteley AS,Burkill PH,Zubkov MVdoi
10.1111/j.1574-6941.2010.00927.xsubject
Has Abstractpub_date
2010-10-01 00:00:00pages
10-8issue
1eissn
0168-6496issn
1574-6941pii
FEM927journal_volume
74pub_type
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