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Purification of Protein-complexes from the Cyanobacterium Synechocystis sp. PCC 6803 Using FLAG-affinity Chromatography

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Preparation of Actinoplanes missouriensis Zoospores and Assay for Their Adherence to Solid Surfaces

Authors: Takeaki Tezuka, Daisuke Nakane, Tomohiro Kimura and Yasuo Ohnishi, date: 12/20/2019, view: 504, Q&A: 0

[Abstract] Spherical zoospores of a rare actinomycete, Actinoplanes missouriensis, adhere to various hydrophobic solid surfaces by means of type IV pili. The purpose of this protocol is to provide detailed descriptions of the preparation of A. missouriensis zoospores and an assay for the adhesion of the zoospores to solid surfaces. This ...

Imaging Cryptococcus spp. Capsule by Differential Interference Contrast Microscopy Using Percoll^®

Authors: Hugo Costa Paes, Stefânia de Oliveira Frazão, Maria Sueli Soares Felipe, Arturo Casadevall and André Moraes Nicola, date: 11/20/2019, view: 661, Q&A: 0

[Abstract] The most important virulence factor in the Cryptococcus genus is the polysaccharide capsule. This genus includes several species that cause life-threatening invasive disease. An increase in capsule thickness is important during fungal infection. The capsule is usually imaged using India ink, and crucial insights on the dynamics of its ...

Purification and HPLC Analysis of Cell Wall Muropeptides from Caulobacter crescentus

Authors: Gabriele Stankeviciute and Eric A. Klein, date: 11/05/2019, view: 790, 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 ...

High Resolution Respirometry in Candida albicans

Authors: Lucian Duvenage, Carol A. Munro and Campbell W. Gourlay, date: 09/05/2019, view: 1198, Q&A: 0

[Abstract] Many Candida species, such as the opportunistic human pathogen Candida albicans, are Crabtree-Negative yeasts and are therefore highly dependent on the energy generated through oxidative phosphorylation. Respiration control is linked to a range of aspects of C. albicans cell physiology that appear to be important for ...

Quantitative Transformation Efficiency Assay for Bacillus subtilis

Authors: Christian L. Loyo and Briana M. Burton, date: 12/05/2018, view: 3121, Q&A: 0

[Abstract] Bacillus subtilis (B. subtilis) is a model Gram-positive organism used to study a variety of physiological states and stress responses, one of which is the development of competence. Competence refers to the physiological state of a cell which allows it to be transformed naturally. Through induction of competence, the efficiency ...

Extraction and Analysis of Bacterial Teichoic Acids

Authors: Kelvin Kho and Timothy C. Meredith, date: 11/05/2018, view: 3364, Q&A: 0

[Abstract] Teichoic acids (TA) are anionic polymers comprised of polyol phosphate repeat units that are abundant in the cell wall of Gram-positive bacteria. Both wall teichoic acid (WTA) and lipoteichoic acid (LTA) play important roles in regulating cell wall remodeling as well as conferring antibiotic resistance. To analyze TA, we describe a polyacrylamide ...

Extraction and Quantification of Polyphosphate (polyP) from Gram-negative Bacteria

Authors: Jan-Ulrik Dahl, Lihan Xie and Ursula Jakob, date: 09/20/2018, view: 2778, Q&A: 0

[Abstract] Polyphosphate (polyP), a universally conserved biomolecule, is composed of up to 1,000 phosphate monomers linked via phosphoanhydride bonds. Reaching levels in bacteria that are in the high nmoles per mg protein range, polyP plays important roles in biofilm formation and colonization, general stress protection and virulence. Various protocols for ...

Selective Isolation of Retroviruses from Extracellular Vesicles by Intact Virion Immunoprecipitation

Authors: Tyler Milston Renner, Kasandra Bélanger and Marc-André Langlois, date: 09/05/2018, view: 2794, Q&A: 0

[Abstract] There exists a wide variety of techniques to isolate and purify viral particles from cell culture supernatants. However, these techniques vary greatly in ease of use, purity, yield and impact on viral structural integrity. Most importantly, it is becoming evident that secreted extracellular vesicles (EVs) co-purify with retroviruses using nearly ...

Quantification of Hydrogen Sulfide and Cysteine Excreted by Bacterial Cells

Authors: Sergey Korshunov and James A. Imlay, date: 05/20/2018, view: 3340, Q&A: 0

[Abstract] Bacteria release cysteine to moderate the size of their intracellular pools. They can also evolve hydrogen sulfide, either through dissimilatory reduction of oxidized forms of sulfur or through the deliberate or inadvertent degradation of intracellular cysteine. These processes can have important consequences upon microbial communities, because ...

Evaluation of the Condition of Respiration and Photosynthesis by Measuring Chlorophyll Fluorescence in Cyanobacteria

Authors: Takako Ogawa and Kintake Sonoike , date: 05/05/2018, view: 3847, Q&A: 0

[Abstract] Chlorophyll fluorescence measurements have been widely used to monitor the condition of photosynthesis. Furthermore, chlorophyll fluorescence from cyanobacteria reflects the condition of respiration, since cyanobacterial photosynthesis shares several components of electron transport chain with respiration. This protocol presents the method to ...

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