scholarly journals Use of green fluorescent protein for detection of cell-specific gene expression and subcellular protein localization during sporulation in Bacillus subtilis

Microbiology ◽  
1996 ◽  
Vol 142 (4) ◽  
pp. 733-740 ◽  
Author(s):  
P. J. Lewis ◽  
J. Errington
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Protein Localization ◽  
Specific Gene ◽  
Specific Gene Expression ◽  
Subcellular Protein Localization ◽  
Green Fluorescent

scholarly journals Benchmarking Various Green Fluorescent Protein Variants in Bacillus subtilis, Streptococcus pneumoniae, and Lactococcus lactis for Live Cell Imaging

2013 ◽  
Vol 79 (20) ◽  
pp. 6481-6490 ◽  
Author(s):  
Wout Overkamp ◽  
Katrin Beilharz ◽  
Ruud Detert Oude Weme ◽  
Ana Solopova ◽  
Harma Karsens ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Green Fluorescent Protein ◽  
Streptococcus Pneumoniae ◽  
Lactococcus Lactis ◽  
Cell Biology ◽  
Fluorescent Protein ◽  
Protein Localization ◽  
Experimental Conditions ◽  
Content Type ◽  
Green Fluorescent

ABSTRACTGreen fluorescent protein (GFP) offers efficient ways of visualizing promoter activity and protein localizationin vivo, and many different variants are currently available to study bacterial cell biology. Which of these variants is best suited for a certain bacterial strain, goal, or experimental condition is not clear. Here, we have designed and constructed two “superfolder” GFPs with codon adaptation specifically forBacillus subtilisandStreptococcus pneumoniaeand have benchmarked them against five other previously available variants of GFP inB. subtilis,S. pneumoniae, andLactococcus lactis, using promoter-gfpfusions. Surprisingly, the best-performing GFP under our experimental conditions inB. subtiliswas the one codon optimized forS. pneumoniaeandvice versa. The data and tools described in this study will be useful for cell biology studies in low-GC-rich Gram-positive bacteria.

scholarly journals Dynamic Patterns of Subcellular Protein Localization during Spore Coat Morphogenesis in Bacillus subtilis

2004 ◽  
Vol 186 (14) ◽  
pp. 4441-4448 ◽  
Author(s):  
Christiaan van Ooij ◽  
Patrick Eichenberger ◽  
Richard Losick
Keyword(s):  
Bacillus Subtilis ◽  
Subcellular Localization ◽  
Fluorescent Protein ◽  
Protein Localization ◽  
Distinct Pattern ◽  
Spore Coat ◽  
Short Axis ◽  
Subcellular Protein Localization ◽  
Associated Proteins ◽  
Green Fluorescent

ABSTRACT Endospores of Bacillus subtilis are encased in a thick, proteinaceous shell known as the coat, which is composed of a large number of different proteins. Here we report the identification of three previously uncharacterized coat-associated proteins, YabP, YheD, and YutH, and their patterns of subcellular localization during the process of sporulation, obtained by using fusions of the proteins to the green fluorescent protein (GFP). YabP-GFP was found to form both a shell and a ring around the center of the forespore across the short axis of the sporangium. YheD-GFP, in contrast, formed two rings around the forespore that were offset from its midpoint, before it eventually redistributed to form a shell around the developing spore. Finally, YutH-GFP initially localized to a focus at one end of the forespore, which then underwent transformation into a ring that was located adjacent to the forespore. Next, the ring became a cap at the mother cell pole of the forespore that eventually spread around the entire developing spore. Thus, each protein exhibited its own distinct pattern of subcellular localization during the course of coat morphogenesis. We concluded that spore coat assembly is a dynamic process involving diverse patterns of protein assembly and localization.

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