Prokaryotes

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    Protocols in Current Issue
    Detection of in vivo Protein Interactions in All Bacterial Compartments by Förster Resonance Energy Transfer with the Superfolder mTurquoise2 ox-mNeongreen FRET Pair
    Authors:  Nils Y. Meiresonne, Elisa Consoli, Laureen M.Y. Mertens and Tanneke den Blaauwen, date: 12/05/2019, view: 151, Q&A: 0
    This protocol was developed to qualitatively and quantitatively detect protein-protein interactions in all compartments of Escherichia coli by Förster Resonance Energy Transfer (FRET) using the Superfolder mTurquoise2 ox-mNeonGreen FRET ...
    Detection of in vivo Protein Interactions in All Bacterial Compartments by Förster Resonance Energy Transfer with the Superfolder mTurquoise2 ox-mNeongreen FRET Pair
    Authors:  Nils Y. Meiresonne, Elisa Consoli, Laureen M.Y. Mertens and Tanneke den Blaauwen, date: 12/05/2019, view: 151, Q&A: 0
    [Abstract] This protocol was developed to qualitatively and quantitatively detect protein-protein interactions in all compartments of Escherichia coli by Förster Resonance Energy Transfer (FRET) using the Superfolder mTurquoise2 ox-mNeonGreen FRET pair (sfTq2ox-mNG). This FRET pair has more than twice the detection range for FRET ...
    Bacterial Synchronized Transfer Assays in Bone Marrow Derived Macrophages
    Authors:  Sedelia R Dominguez, Tom Kawula and Shaun P Steele, date: 11/20/2019, view: 307, Q&A: 0
    [Abstract] Merocytophagy (“mero”, Greek for partial; “cytophagy” for cell eating) is a process by which cells acquire microbes and cytosolic material through phagocytosis of a small portion of neighboring cells upon cell-cell contact. Cell-cell contact dependent transfer events can be assessed through co-incubation of differently labeled cells. With these ...
    Assessing Different Ways of Bacillus subtilis Spreading over Abiotic Surfaces
    Authors:  Marco Bartolini and Roberto Grau, date: 11/20/2019, view: 326, Q&A: 0
    [Abstract] Surface-associate motility on biotic and abiotic environments is a key mechanism used by the model bacterium Bacillus subtilis and its closest relatives (i.e., B. amyloliquefaciens, B. thuringiensis, B. cereus, B. pumilus) for surface colonization and spreading across surfaces. The study of ...
    Super-resolution Microscopy at Cryogenic Temperatures Using Solid Immersion Lenses
    [Abstract] Our mechanistic understanding of cell function depends on imaging biological processes in cells with molecular resolution. Super-resolution fluorescence microscopy plays a crucial role by reporting cellular ultrastructure with 20-30 nm resolution. However, this resolution is insufficient to image macro-molecular machinery at work. A path to ...
    Purification and HPLC Analysis of Cell Wall Muropeptides from Caulobacter crescentus
    Authors:  Gabriele Stankeviciute and Eric A. Klein, date: 11/05/2019, view: 270, Q&A: 0
    [Abstract] The peptidoglycan sacculus, or cell wall, is what defines bacterial cell shape. Cell wall composition can be best characterized at the molecular level by digesting the peptidoglycan murein polymer into its muropeptide subunits and quantifying the abundance of muropeptides using high-pressure liquid chromatography. Certain features of the cell wall ...
    Probabilistic Models for Predicting Mutational Routes to New Adaptive Phenotypes
    Authors:  Eric Libby and Peter A. Lind, date: 10/20/2019, view: 531, Q&A: 0
    [Abstract] Understanding the translation of genetic variation to phenotypic variation is a fundamental problem in genetics and evolutionary biology. The introduction of new genetic variation through mutation can lead to new adaptive phenotypes, but the complexity of the genotype-to-phenotype map makes it challenging to predict the phenotypic effects of ...
    Supported Cell Membrane Sheets to Monitor Protein Assembly
    Authors:  Simon Erlendsson, Thor Seneca Thorsen and Kenneth Lindegaard Madsen, date: 09/20/2019, view: 462, Q&A: 0
    [Abstract] Studying protein-protein and protein-lipid interactions in their native environment is highly desirable, yet, the heterogeneity and complexity of cellular systems limits the repertoire of experimental methods available. In cells, interactions are often taking place in confined microenvironments where factors such as avidity, hindered diffusion, ...
    A PhoA-STII Based Method for Efficient Extracellular Secretion and Purification of Fab from Escherichia coli
    Authors:  Ziyan Wang, Yang Gao, Manyu Luo, Cedric Cagliero, Hua Jiang, Yueqing Xie, Jianwei Zhu and Huili Lu, date: 09/20/2019, view: 432, Q&A: 0
    [Abstract] In comparison with full-length IgGs, antigen binding fragments (Fabs) are smaller in size and do not require the complexed post-translational modification. Therefore, Fab can be cost-effectively produced using an Escherichia coli (E. coli) expression system. However, the disulfide-bonds containing exogenous protein, including ...
    Cryo-transmission Electron Microscopy of Outer-inner Membrane Vesicles Naturally Secreted by Gram-negative Pathogenic Bacteria
    [Abstract] A protocol was developed to visualize and analyze the structure of membrane vesicles (MVs) from Gram-negative bacteria. It is now accepted that these micrometric spherical vesicles are commonly produced by cells from all three domains of life, so the protocol could be useful in the study of vesicles produced by eukaryotes and archaea as well as ...
    Measurement of the Length of the Integrated Donor DNA during Bacillus subtilis Natural Chromosomal Transformation
    Authors:  Ester Serrano and Begoña Carrasco, date: 08/20/2019, view: 485, Q&A: 0
    [Abstract] For natural transformation to occur, bacterial cells must first develop a programmed physiological state called competence. Competence in Bacillus subtilis, which occurs only in a fraction of cells, is a transient stress response that allows cells to take up DNA from the environment. During natural chromosomal transformation, the ...



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