2015
DOI: 10.1128/jb.02221-14
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Bright Fluorescent Streptococcus pneumoniae for Live-Cell Imaging of Host-Pathogen Interactions

Abstract: c Streptococcus pneumoniae is a common nasopharyngeal resident in healthy people but, at the same time, one of the major causes of infectious diseases such as pneumonia, meningitis, and sepsis. The shift from commensal to pathogen and its interaction with host cells are poorly understood. One of the major limitations for research on pneumococcal-host interactions is the lack of suitable tools for live-cell imaging. To address this issue, we developed a generally applicable strategy to create genetically stable… Show more

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Cited by 75 publications
(94 citation statements)
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“…A) A phase contrast image of VL2706. B) ComEA fusions produces strong foci around midcell as well as at the poles of the cell as previously shown (31), C) HlpA fusions produce distinct green signal that localizes at the chromosome(32,33), and D) SPV_1159 fusions produce distinct membrane associated signals in the bacterial cells. E) An overlay of the three simultaneously expressed fusion proteins and a yellow outline of F) containing an expanded view of the image.…”
supporting
confidence: 69%
See 1 more Smart Citation
“…A) A phase contrast image of VL2706. B) ComEA fusions produces strong foci around midcell as well as at the poles of the cell as previously shown (31), C) HlpA fusions produce distinct green signal that localizes at the chromosome(32,33), and D) SPV_1159 fusions produce distinct membrane associated signals in the bacterial cells. E) An overlay of the three simultaneously expressed fusion proteins and a yellow outline of F) containing an expanded view of the image.…”
supporting
confidence: 69%
“…We selected ComEA as this is a protein involved in DNA uptake and was previously shown to demonstrate a septal localization (31). HlpA (HU) was chosen as it binds to the nucleoid and is a good marker for the chromosome (32,33). SPV_1159 was selected as it is a predicted small membrane protein with two transmembrane domains with both the N-and C-terminus inside the cytoplasm and was previously shown to demonstrate homogenous membrane localization in our laboratory (Gallay and Veening, unpublished).…”
Section: Simultaneous Integration and Expression Of Three New Fluoresmentioning
confidence: 99%
“…Live-cell imaging of strains showing fluorescence or bioluminescence has been proposed as an alternative to FISH. For example, highly fluorescent S. pneumoniae has been constructed by using a superfolder green fluorescent protein as a reporter (34). Nevertheless, when we have used these constructs to produce NE S. pneumoniae and NE S. pneumoniae-NT H. influenzae in vitro biofilms, significant differences in fluorescence among individual bacteria were noted (unpublished observations).…”
Section: Discussionmentioning
confidence: 99%
“…Despite the differences observed with NCTC 7466, D39V is genetically stable under laboratory conditions, as both the hsdS locus and the ter configuration are identical in all analyzed derivative strains (not shown). Additionally and importantly, D39V is virulent in multiple infection models (78)(79)(80)(81), making it an ideal, stable genetic workhorse for pneumococcal research. Therefore, the strain will be made available to the community through the PHE National…”
Section: Discussionmentioning
confidence: 99%