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Production of endo-N in uninduced and induced EV36 harbouring pRep4 and pEndo-N

Authors: Steenbergen, Susan M; Vimr, Eric R;

Production of endo-N in uninduced and induced EV36 harbouring pRep4 and pEndo-N

Abstract

Bacteria (20 ml) were grown to early stationary phase with appropriate drugs and pelleted by centrifugation. The pellets were washed in phosphate-buffered saline (PBS), re-suspended in PBS to final volume of 0.5 ml and disrupted by sonication. The sonicate was centrifuged for 15 min at 35 000 and 25 μl amounts of supernatants either loaded with an equal volume of PBS into wells of a radial immunodiffusion plate containing 10% (v/v) anti-polysialic acid antibody, or pre-incubated for 15 min at 37°C with an equal volume of induced or uninduced extract from EV36 harbouring pRep4 and pEndo-N prior to loading. Wells 1–4 contained extracts from EV36, EV93, EV36 pRep4 pEndo-N and induced EV36 pRep4 pEndo-N respectively. Wells 5 and 6 contained EV93 extracts pre-incubated with uninduced or induced extracts of EV36 harbouring pRep4 and pEndo-N respectively. Precipitin halos indicate diffusion of PSA and reaction with the antibody. The photograph was taken after overnight incubation at 32°C and 1 day at 4°C.Copyright information:Taken from "Biosynthesis of the K1 group 2 polysialic acid capsule occurs within a protected cytoplasmic compartment"Molecular Microbiology 2008;68(5):1252-1267.Published online Jan 2008PMCID:PMC2408645.© 2008 The Authors. Journal compilation © 2008 Blackwell Publishing Ltd

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  • citations
    This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    0
    popularity
    This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
    Average
    influence
    This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    Average
    impulse
    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
    Average
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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
0
Average
Average
Average