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Generation of Giant Unilamellar Vesicles (GUVs) Using Polyacrylamide Gels

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Bacterial Conjugation Protocol for Ruminant Mycoplasmas

Authors: Eveline Sagné, Christine Citti and Emilie Dordet-Frisoni, date: 01/20/2021, view: 412, Q&A: 0

[Abstract]

In Mycoplasma agalactiae, two simultaneous processes of DNA transfer have been described that require direct cell-to-cell contact and are similar to conjugation. One involves the self-transmission of an integrative conjugative element (ICE) while the second concerns the horizontal transfer of large and small fragments of chromosomal DNA. Here, we

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Charging State Analysis of Transfer RNA from an α-proteobacterium

Authors: Liang Yin and Caroline S. Harwood, date: 12/05/2020, view: 504, Q&A: 0

[Abstract] Transfer RNA (tRNA) is an essential link between the genetic code and proteins. During the process of translation, tRNA is charged with its cognate amino acid and delivers it to the ribosome, thus serving as a substrate of protein synthesis. To analyze the charging state of a particular tRNA, total RNA is purified and analyzed on an acid-urea gel. ...

Ribosome Purification from an α-proteobacterium and rRNA Analysis by Northern Blot

Authors: Liang Yin and Caroline S. Harwood, date: 12/05/2020, view: 486, Q&A: 0

[Abstract] Ribosomes are an integral part of cellular life. They are complex molecular machines consisting of multiple ribosomal proteins and RNAs. To study different aspects of ribosome composition, many methods have been developed over the decades. Here, we describe how to purify ribosomes from the α-proteobacterium Rhodopseudomonas palustris ...

A High-throughput Interbacterial Competition Platform

Authors: Hsiao-Han Lin and Erh-Min Lai, date: 09/05/2020, view: 1300, Q&A: 0

[Abstract] Contact-dependent interbacterial competition is a common strategy used by bacteria to fight for their ecological niches. Interbacterial competition is monitored by a competition assay involving co-culturing the attacker and the recipient bacterial cells on agar, followed by recovery of the surviving recipient cells. Conventional interbacterial ...

High-level Production of Recombinant Membrane Proteins Using the Engineered Escherichia coli Strains SuptoxD and SuptoxR

Authors: Myrsini Michou, Dafni C. Delivoria and Georgios Skretas, date: 08/05/2020, view: 1509, Q&A: 0

[Abstract] We have previously described the development of two specialized Escherichia coli strains for high-level recombinant membrane protein (MP) production. These engineered strains, termed SuptoxD and SuptoxR, are capable of suppressing the cytotoxicity caused by MP overexpression and of producing greatly enhanced MP yields. Here, we present a ...

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: 1481, 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: 1500, 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: 2072, 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: 2067, 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: 4459, 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 ...

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