• 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

Determination of Intra- and Extracellular Glucose in Mycelium of Fusarium oxysporum Authors:  Carmen Ruiz-Roldan and M. Isabel G. Roncero, date: 07/20/2016, view: 5553, Q&A: 0
To study alterations in the metabolism and/or in the transport of glucose during Fusarium oxysporum vegetative growth, we determined intracellular glucose levels in different fungal strains, as well as the amount of glucose remaining in the supernatants after growth in synthetic medium (SM) supplemented with either 0.05 or 2.5% glucose. ...
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Determination of Intracellular ATP Levels in Mycelium of Fusarium oxysporum Authors:  Carmen Ruiz-Roldan and M. Isabel G. Roncero, date: 07/20/2016, view: 4913, Q&A: 0
Glycolysis provides metabolites for energy production via oxidative phosphorylation during vegetative growth of Fusarium oxysporum. Therefore, determination of intracellular ATP levels might be of valuable help to analyze regulation of glycolysis/gluconeogenesis pathways. The protocol described here can be applied to other filamentous ...
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Expression, Purification and Crystallization of the Herpesvirus Nuclear Egress Complex (NEC) Authors:  Janna M. Bigalke and Ekaterina E. Heldwein, date: 07/20/2016, view: 4339, Q&A: 0
The protocol describes the production and crystallization of the soluble form of the nuclear egress complex (NEC) from Herpes simplex virus 1 and Pseudorabies virus. The NEC is a heterodimer that consists of conserved proteins UL31 and UL34. NEC oligomerization deforms the inner nuclear membrane around the capsid in infected cells, thereby ...
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Investigating the Assembly Status of the Plastid Encoded Polymerase Using BN-PAGE and Sucrose Gradient Centrifugation Author:  Jeannette Pfalz, date: 07/20/2016, view: 4615, Q&A: 0
The plastid encoded polymerase (PEP) represents a major transcription machinery in mature chloroplasts (Liere et al., 2011; Zhelyazkova et al., 2012). The proper assembly of this multi-subunit complex is important for plant growth and development (Pfalz and Pfannschmidt, 2013). The PEP polymerase can be purified from soluble and ...
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Ubiquitination Assay for Mammalian Cells Authors:  Yang Peng, Edward Wang, Guang Peng and Shiaw-Yih Lin, date: 07/20/2016, view: 11115, Q&A: 2
Ubiquitin is an 8.5 kDa protein that can be activated and conjugated by ubiquitin-activating enzyme E1 and ubiquitin-conjugating enzyme E2, respectively. Ubiquitin E3 ligases then recognize protein substrates, and then transfer the ubiquitin from E2 to the targeted protein. This biological process is called ubiquitination, and it is an important ...
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Developmental Biology

Differentiation of Human Embryonic Stem Cells into Cone Photoreceptors Authors:  Anthony Flamier, Andrea Barabino and Gilbert Bernier, date: 07/20/2016, view: 4957, Q&A: 1
Photoreceptors are specialized retinal neurons able to respond to light in order to generate visual information. Among photoreceptors, cones are involved in colors discrimination and high-resolution central vision and are selectively depleted in macular degenerations and cone dystrophies. A possible therapeutic solution for these disorders is to ...
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Immunology

House Dust Mite Extract and Cytokine Instillation of Mouse Airways and Subsequent Cellular Analysis Authors:  Chris Tibbitt and Jonathan M. Coquet, date: 07/20/2016, view: 6027, Q&A: 0
Asthma is a complex disease of the airways primarily mediated by T helper 2 cells and innate lymphoid type 2 cells (Licona et al.,2013). Mice do not develop spontaneous asthma and therefore models have been developed for the assessment of key processes that underlie human pathology (Nial et al.,2008). Exposure to House Dust Mite ...
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Microbiology

A Bioassay Protocol for Quorum Sensing Studies Using Vibrio campbellii Authors:  Alberto J. Martín-Rodríguez and José J. Fernández, date: 07/20/2016, view: 5698, Q&A: 0
Quorum Sensing (QS), or bacterial cell-to-cell communication, is a finely-tuned mechanism that regulates gene expression on a population density-dependent manner through the production, secretion and reception of extracellular signaling molecules termed autoinducers (AIs). Given that QS plays an important role in bacterial biofilm formation and ...
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Reporter Assay for Semen-mediated Enhancement of HIV-1 Infection Authors:  Janis A. Müller and Jan Münch, date: 07/20/2016, view: 5529, Q&A: 0
Semen contains amyloid fibrils that enhance HIV-1 infection (Münch et al., 2007; Kim et al., 2010; Roan et al., 2011; Arnold et al., 2012; Usmani et al., 2014; Roan et al., 2014). Positively charged semen amyloids capture negatively charged viral particles and increase their attachment rates to ...
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Extraction and Quantification of Polyphosphate in the Budding Yeast Saccharomyces cerevisiae Authors:  David Canadell, Samuel Bru, Josep Clotet and Joaquín Ariño, date: 07/20/2016, view: 6235, Q&A: 0
Inorganic polyphosphate (polyP) is a linear polymer present in both prokaryotic and eukaryotic organisms and made from three to hundreds of orthophosphate residues linked by phosphoanhydride bonds. The biological role of this molecule goes beyond serving as Pi store or energy source to replace ATP. For instance, in yeast polyP levels have been ...
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Fluorescent Detection of Intracellular Nitric Oxide in Staphylococcus aureus Authors:  April M. Lewis, Silvia S. Matzdorf and Kelly C. Rice, date: 07/20/2016, view: 4736, Q&A: 0
Nitric Oxide (NO) is a highly-reactive radical gas that can modify a variety of cellular targets in both eukaryotes and bacteria. NO is produced endogenously by a wide variety of organisms: For example, as a cell-signaling molecule in mammals and bacteria via nitric oxide synthase (NOS) enzymes, and as a product of denitrification. As such, it is ...
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Separation of Free and Bound cAMP in Mycobacteria Authors:  Arka Banerjee and Sandhya S. Visweswariah, date: 07/20/2016, view: 4210, Q&A: 0
Mycobacterial genomes encode a plethora of genes that are involved in the synthesis, utilization and degradation of cAMP. The genome of M. tuberculosis H37Rv, for example, encodes 16 adenylyl cyclases and 10 genes harbouring the cyclic nucleotide-binding (CNB) domain (Shenoy and Visweswariah, 2006). Cyclic AMP is efficiently secreted by ...
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Neuroscience

Mouse Subependymal Zone Explants Cultured on Primary Astrocytes Authors:  Ionut Dumitru, Hannah Monyer and Julieta Alfonso, date: 07/20/2016, view: 4448, Q&A: 0
Neuroblast migration is a major component in the process of neuronal circuit assembly. In the rodent postnatal brain, the subependymal zone (SEZ) is the largest neurogenic niche where new neurons are born throughout life. These cells migrate several millimeters following a specific pathway called the rostral migratory stream (RMS) towards the ...
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Plant Science

Isolation of Flavonoids from Piper delineatum Leaves by Chromatographic Techniques Authors:  Isabel L. Bazzocchi, Juan C. Ticona, Ignacio A. Jiménez and Ninoska Flores, date: 07/20/2016, view: 9434, Q&A: 0
The genus Piper (Piperaceae) is widely distributed in the tropical and subtropical regions of the world, and species belonging to this genus are included in the Ayurvedic system of medicine and in folklore medicine of Latin America. Phytochemical investigations of Piper species have led to the isolation of several classes of ...
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Establishment of a Fusarium graminearum Infection Model in Arabidopsis thaliana Leaves and Floral Tissues Authors:  Vamsi J Nalam, Sujon Sarowar and Jyoti Shah, date: 07/20/2016, view: 6364, Q&A: 0
Fusarium graminearum (Fg) is the causal agent of Fusarium head blight disease of wheat (Triticum aestivum), oats (Avena sativa) and barley (Hordeum vulgare), which targets the floral tissues and thereby adversely impacts grain yield and quality. Mycotoxins produced by F. graminearum further limit the ...
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