• Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Biochemistry

Nanofluidic Proteomic Immunoassay Authors:  Huifang Guo, Philip L Lorenzi and Zhiyong Ding, date: 07/20/2015, view: 5262, Q&A: 0
Nanofluidic proteomic immunoassay (NIA), consisting of isoelectric focusing followed by sensitive chemiluminescence detection, has the potential to quantitatively characterize protein post-translational modifications that cause shifts in isoelectric point (pI). This protocol details the NIA analysis of protein phosphorylation using AKT as an ...
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Micro-scale NMR Experiments for Monitoring the Optimization of Membrane Protein Solutions for Structural Biology Authors:  Reto Horst and Kurt Wüthrich, date: 07/20/2015, view: 4927, Q&A: 0
Reconstitution of integral membrane proteins (IMP) in aqueous solutions of detergent micelles has been extensively used in structural biology, using either X-ray crystallography or NMR in solution. Further progress could be achieved by establishing a rational basis for the selection of detergent and buffer conditions, since the stringent ...
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Cell Biology

A Simple Method to Generate Gene Knockout Clones in Human Cells Using Transcription Activator-Like Effector Nuclease (TALEN) Author:  Christy Hammack, date: 07/20/2015, view: 6487, Q&A: 0
Transcription activator-like effectors (TALEs) are naturally occurring proteins secreted by the plant pathogen, Xanthomonas, and fused to the Fok1 endonuclease to generate TALE nucleases (TALENs). TALEN pairs bind to specific DNA sequences initiating Fok1 dimerization and double-stand cleavage of DNA within the TALEN target site. This ...
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Microbiology

Genetic Transformation of Candida glabrata by Electroporation Authors:  Fabian Istel, Tobias Schwarzmüller, Michael Tscherner and Karl Kuchler, date: 07/20/2015, view: 5545, Q&A: 0
Here, we report a method for the transformation by electroporation of the human fungal pathogen Candida glabrata (C. glabrata). The protocol can be used for transformations in single well or in 96-well microtiter plates. It has been extensively used to generate a genome-scale gene deletion library using the C. glabrata ...
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Genetic Transformation of Candida glabrata by Heat Shock Authors:  Fabian Istel, Tobias Schwarzmüller, Michael Tscherner and Karl Kuchler, date: 07/20/2015, view: 6244, Q&A: 0
Here, we report a method for the transformation of Candida glabrata using a heat shock method. The protocol can be used for transformations in single well or in 96-well scale. It has been employed as an alternative method to the electroporation protocol to construct a genome-scale gene deletion collection in the human fungal pathogen ...
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Large-scale Phenotypic Profiling of Gene Deletion Mutants in Candida glabrata Authors:  Fabian Istel, Tobias Schwarzmüller, Michael Tscherner and Karl Kuchler, date: 07/20/2015, view: 7140, Q&A: 0
Here, we describe a method enabling the phenotypic profiling of genome-scale deletion collections of fungal mutants to detect phenotypes for various stress conditions. These stress conditions include among many others antifungal drug susceptibility, temperature-induced and osmotic as well as heavy metal or oxidative stress. The protocol was ...
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Generation of Mammalian Host-adapted Leptospira interrogans by Cultivation in Peritoneal Dialysis Membrane Chamber Implantation in Rats Authors:  André Alex Grassmann, Alan John Alexander McBride, Jarlath E. Nally and Melissa J. Caimano, date: 07/20/2015, view: 5896, Q&A: 0
Leptospira interrogans can infect a myriad of mammalian hosts, including humans (Bharti et al., 2003; Ko et al., 2009). Following acquisition by a suitable host, leptospires disseminate via the bloodstream to multiple tissues, including the kidneys, where they adhere to and colonize the proximal convoluted renal ...
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Molecular Biology

In vitro DNA Protection Assay Using Oxidative Stress Authors:  Yuri Ushijima, Ryosuke L. Ohniwa and Kazuya Morikawa, date: 07/20/2015, view: 6189, Q&A: 0
A wide range of stresses such as oxidative stress, acid, alkaline, UV, and metal can damage DNA. Here, we describe a protocol to measure the DNA nicking damage by Fenton reaction-mediated oxidative stress. Fenton reaction (Fe2+ + H2O2 → Fe3+ + OH- + ∙OH) produces the highly deleterious ...
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Neuroscience

A Rat Model of Intracerebral Hemorrhage Induced by Collagenase IV Authors:  Qin Lu, LiJie Huang and QiChuan ZhuGe, date: 07/20/2015, view: 7745, Q&A: 0
Stroke, the second leading cause of death worldwide (Ingall, 2004), is one of the major causes of morbidity and mortality. Intracerebral hemorrhage (ICH), a lethal type of stroke, accounts for 20% of all strokes (Qureshi et al., 2001), and occurs in about 50-60% of Asians (Inagawa, 2002). In order to understand the disease process, three ...
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Plant Science

A Phosphopeptide Purification Protocol for the Moss Physcomitrella paten Authors:  Xiaoqin Wang and Yikun He, date: 07/20/2015, view: 5745, Q&A: 0
Protein phosphorylation is one of the most common post-translational modifications in eukaryotic cells and plays a critical role in a vast array of cellular processes. Efficient methods of protein extraction and phosphopeptide purification are required to ensure the detection of high quality of proteins. In our hands, phenol extraction of proteins ...
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Chlorophyll Fluorescence Measurements in Arabidopsis Wild-type and Photosystem II Mutant Leaves Authors:  Iris Steinberger, Felix Egidi and Anja Schneider, date: 07/20/2015, view: 7126, Q&A: 0
Chlorophyll fluorescence measurement is a widely used technique to determine photosynthetic performance. Light energy absorbed by a chlorophyll molecule can be dissipated by driving photochemical energy conversion, as heat in non-photochemical quenching processes, or it is re-emitted as fluorescence. The loss of light energy as chlorophyll ...
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Circular RT-PCR Assay Using Arabidopsis Samples Authors:  Runlai Hang, Xian Deng, Chunyan Liu, Beixin Mo and Xiaofeng Cao, date: 07/20/2015, view: 10642, Q&A: 0
Post-transcriptional processing is critical for RNA biogenesis, in which conventional functional RNA transcripts are generated, such as messenger RNAs (mRNAs), transfer RNAs (tRNAs) and ribosomal RNAs (rRNAs) for translation as well as emerging non-coding RNAs with known or unknown regulatory functions. To determine the precise termini of an RNA ...
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Atomic Force Microscopy (AFM) Analysis of Cell Wall Structural Glycoproteins in vitro Authors:  Yuning Chen, Liwei Chen, Marcia J. Kieliszewsk and Maura C. Cannon, date: 07/20/2015, view: 5393, Q&A: 0
Hydroxyproline-rich glycoproteins (HRGPs) are major protein components in dicot primary cell walls and generally account for more than 10% of the wall dry weight. As essential members of the HRGP superfamily, extensins (EXTs) presumably function in the cell wall by assembling into positively charged protein scaffolds (Cannon et al., 2008) ...
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An Evaluation of Cellulose Degradation Affected by Dutch Elm Disease Authors:  Jaroslav Ďurkovič, František Kačík and Dušan Olčák, date: 07/20/2015, view: 5684, Q&A: 0
The pathogenic fungus Ophiostoma novo-ulmi spreads within the secondary xylem vessels of infected elm trees, causing the formation of vessel plugs due to tyloses and gels, which ultimately result in Dutch elm disease. Foliage discoloration, wilting and falling from the tree are typical external leaf symptoms of the disease followed by the ...
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Phakopsora pachyrhizi Infection Bioassay in Detached Soybean Transgenic Leaves for Candidate Gene Validation Authors:  Beatriz Wiebke-Strohm, Ciliana Rechenmacher, Luisa Abruzzi de Oliveira, Cláudia Vieira Godoy and Maria Helena Bodanese Zanettini, date: 07/20/2015, view: 5822, Q&A: 0
Soybean (Glycine max) is one of the most important crops in the world. Phakopsora pachyrhizi is a plant pathogenic basidiomycete fungus that infects soybean, causing Asian Soybean Rust (ASR) disease and affecting production. Here, we describe how to prepare the plant material and the uredospore suspension (from spores harvested ...
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