Metaproteomic and metagenomic analyses of defined oceanic microbial populations using microwave cell fixation and flow cytometric sorting.

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 Ecol

authors

Mary I,Oliver A,Skipp P,Holland R,Topping J,Tarran G,Scanlan DJ,O'Connor CD,Whiteley AS,Burkill PH,Zubkov MV

doi

10.1111/j.1574-6941.2010.00927.x

subject

Has Abstract

pub_date

2010-10-01 00:00:00

pages

10-8

issue

1

eissn

0168-6496

issn

1574-6941

pii

FEM927

journal_volume

74

pub_type

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