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Generation of Giant Unilamellar Vesicles (GUVs) Using Polyacrylamide Gels

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Analysis of Gram-negative Bacteria Peptidoglycan by Ultra-performance Liquid Chromatography

Authors: Laura Alvarez, Baptiste Cordier, Sven van Teeffelen and Felipe Cava, date: 10/05/2020, view: 838, Q&A: 0

[Abstract] Bacteria are surrounded by a protective peptidoglycan cell wall. Provided that this structure and the enzymes involved are the preferred target for our most successful antibiotics, determining its structural and chemical complexity is of the highest interest. Traditionally, high-performance liquid chromatography (HPLC) analyses have been ...

Determination of the Cellular Ion Concentration in Saccharomyces cerevisiae Using ICP-AES

Authors: Louise Thines, Anne Iserentant and Pierre Morsomme, date: 08/20/2020, view: 917, Q&A: 0

[Abstract] The yeast Saccharomyces cerevisiae has been perceived over decades as a highly valuable model organism for the investigation of ion homeostasis. Indeed, many of the genes and biological systems that function in yeast ion homeostasis are conserved throughout unicellular eukaryotes to humans. In this context, measurement of the yeast ...

Quantification of Nitric Oxide and Reactive Oxygen Species in Leishmania-infected J774A.1 Macrophages as a Response to the in vitro Treatment with a Natural Product Compound

Authors: Kalliopi Karampetsou, Olga S. Koutsoni and Eleni Dotsika, date: 12/05/2019, view: 1590, Q&A: 0

[Abstract] Leishmaniasis is a parasitic disease caused by the obligatory intracellular protozoa Leishmania spp. Current therapeutic options are limited and thus, drug discovery against leishmaniasis is very important. Nevertheless, there is a great difficulty to develop therapeutic strategies against the disease because the parasite deploys various ...

Intracellular cAMP Measurements in Candida albicans Biofilms

Authors: Liuliu Jiang, Shengyan Chen, Kairui Sun, Peng Zhou and Xin Wei, date: 12/05/2019, view: 1687, Q&A: 0

[Abstract] Candida albicans is the most common cause of fungal infections worldwide. Infection by C. albicans is closely associated with its ability to form a biofilm, closely packed communities of cells attached to the surfaces of human tissues and implanted devices, in or on the host. When tested for susceptibility to antifungals, such as ...

Detection of D-glutamate Production from the Dual Function Enzyme, 4-amino-4-deoxychorismate Lyase/D-amino Acid Transaminase, in Mycobacterium smegmatis

Authors: Helen K. Opel-Reading, Roman Mortuza and Kurt L. Krause, date: 01/05/2019, view: 3260, Q&A: 0

[Abstract] D-amino acid transaminase (D-AAT) is able to synthesize both D-glutamate and D-alanine, according to the following reaction: D-alanine + α-ketoglutarate ⇌ D-glutamate + pyruvate. These two D-amino acids are essential components of the peptidoglycan layer of bacteria. In our recently published work, MSMEG_5795 from Mycobacterium smegmatis ...

Extraction and Analysis of Bacterial Teichoic Acids

Authors: Kelvin Kho and Timothy C. Meredith, date: 11/05/2018, view: 4978, 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: 3722, 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 ...

Determination of Polyhydroxybutyrate (PHB) Content in Ralstonia eutropha Using Gas Chromatography and Nile Red Staining

Authors: Janina R. Juengert, Stephanie Bresan and Dieter Jendrossek, date: 03/05/2018, view: 10307, Q&A: 0

[Abstract] Ralstonia eutropha H16 produces and mobilizes (re-utilizes) intracellular polyhydroxybutyrate (PHB) granules during growth. This protocol describes the visualization of intracellular Nile red stained PHB granules and the quantification of PHB by gas chromatography. Our first method describes how to analyze PHB granules by fluorescence ...

Identification and Quantification of Secondary Metabolites by LC-MS from Plant-associated Pseudomonas aurantiaca and Pseudomonas chlororaphis

Authors: Izzah Shahid, Muhammad Rizwan and Samina Mehnaz, date: 01/20/2018, view: 8019, Q&A: 0

[Abstract] Increased antibiotic resistance of plants and human pathogens and continuous use of chemical fertilizers has pushed microbiologists to explore new microbial sources as potential antagonists. In this study, eight strains of Pseudomonas aurantiaca and Pseudomonas chlororaphis, have been isolated from different plant sources and ...

Determination of Mn Concentrations in Synechocystis sp. PCC6803 Using ICP-MS

Authors: Fabian Brandenburg, Hanan Schoffman, Nir Keren and Marion Eisenhut, date: 12/05/2017, view: 5215, Q&A: 0

[Abstract] Manganese (Mn) is an essential micronutrient for all photoautotrophic organisms. Two distinct pools of Mn have been identified in the cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis), with 80% of the Mn residing in the periplasm and 20% in cytoplasm and thylakoid lumen (Keren et al., 2002). In this protocol, we ...

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