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Past Issues in 2014

Volume: 4 Issue: 17

Biochemistry

Analysis of L- and D-Amino Acids Using UPLC Authors: Yuta Mutaguchi and Toshihisa Ohshima, date: 09/05/2014, view: 8333, Q&A: 0

With the exception of glycine, α-amino acids are optically active, and two optical isomers (L- and D-) of each amino acid can be formed. Recent developments of analytical techniques have revealed that several free D-amino acids such as D-aspartate, D-serine and D-alanine exist in many kinds of organism including human and have biologically ...

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Cell Biology

Dye-uptake Experiment through Connexin Hemichannels Authors: Andrea Puhar and Philippe J. Sansonetti, date: 09/05/2014, view: 7394, Q&A: 0

Connexins (Cxs) are integral membrane proteins of vertebrates that associate to form hexameric transmembrane channels, named hemichannels. Twenty-one Cx types have been described, which are named according to their molecular weight. Cxs are expressed in many cell types, e.g. epithelial cells, astrocytes and immune cells. Hemichannels ...

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Microbiology

Metabolite and Fatty Acid Analysis of Yeast Cells and Culture Supernatants Authors: Liwei Chen and Wei Ning Chen, date: 09/05/2014, view: 10945, Q&A: 0

Metabolite and fatty acid analysis play important roles in evaluating the metabolic state of microorganisms. To examine the growth state and metabolism response of cells to environmental stress or genetic modification, intracellular and extracellular metabolites including fatty acids are usually analyzed to help understand the cellular biochemical ...

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Induction of Connexin-hemichannel Opening Authors: Andrea Puhar and Philippe J. Sansonetti, date: 09/05/2014, view: 5260, Q&A: 0

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Purification of the GfsA-3x FLAG Protein Expressed in Aspergillus nidulans Authors: Takuji Oka, Yukako Katafuchi, Kohsai Fukuda, Keisuke Ekino, Masatoshi Goto and Yoshiyuki Nomura, date: 09/05/2014, view: 5428, Q&A: 0

GfsA is a fungal β-galactofuranosyltransferase involved in the biosynthesis of O-glycan. To investigate the enzymatic functions of GfsA, we attempted to obtain a recombinant protein of this enzyme from two heterologous host organisms. However, GfsA could not be expressed as a recombinant protein in either Escherichia coli (E. ...

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Determination of D-galactofuranose Content of Galactomannoproteins in Aspergillus nidulans Authors: Takuji Oka, Yukako Katafuchi, Kohsai Fukuda, Keisuke Ekino, Masatoshi Goto and Yoshiyuki Nomura, date: 09/05/2014, view: 4963, Q&A: 0

Galactofuranose (Galf) is a component of several polysaccharides and glycoconjugates in certain species of filamentous fungi. Galf residues are frequently found in Aspergillus glycoproteins, including N-glycans and O-mannose glycans that modify many cell wall proteins and extracellular enzymes. It is ...

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Iodine Staining of Escherichia coli Expressing Genes Involved in the Synthesis of Bacterial Glycogen Authors: Ana M. Demonte, Matías D. Asención Diez, Sergio A. Guerrero, Miguel A. Ballicora and Alberto A. Iglesias, date: 09/05/2014, view: 7182, Q&A: 0

The presence of intracellular glycogen can be detected by the following iodine staining technique. Cells with glycogen stain dark brown, whereas in its absence they remain with a pale yellowish color. It is hypothesized that iodine atoms fit into helical coils formed by the α-polyglucan to form a coloured glycogen-iodine complex. Here, we have ...

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Evolution of Escherichia coli to Macrophage Cell Line Authors: Migla Miskinyte and Isabel Gordo, date: 09/05/2014, view: 6593, Q&A: 0

The genomes of species of Escherichia coli (E. coli) show an extraordinary amount of diversity, which include commensal strains and strains belonging to different pathovars. Many strains of E. coli, which can cause mild or severe pathologies in humans, have a commensal ancestor. Understanding the evolutionary changes ...

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Fitness Measurements of Evolved Esherichia coli Authors: Migla Miskinyte and Isabel Gordo, date: 09/05/2014, view: 7217, Q&A: 0

Bacteria can adapt very rapidly to novel selective pressures. In the transition from commensalism to pathogenicity bacteria have to face and adapt to the host immune system. Specifically, the antagonistic interaction imposed by one of the first line of defense of innate immunity cells, macrophages, on commensal bacteria, such as Escherichia coli ...

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Plant Science

Targeted Gene Mutation in Rice Using a CRISPR-Cas9 System Authors: Kabin Xie, Bastian Minkenberg and Yinong Yang, date: 09/05/2014, view: 21273, Q&A: 0

RNA-guided genome editing (RGE) using bacterial type II cluster regularly interspaced short palindromic repeats (CRISPR)–associated nuclease (Cas) has emerged as a simple and versatile tool for genome editing in many organisms including plant and crop species. In RGE based on the Streptococcus pyogenes CRISPR-Cas9 system, the Cas9 ...

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Isolation of Mitochondria from Potato Tubers Authors: Jesper F. Havelund, Fernanda Salvato, Mingjie Chen, R.S.P. Rao, Adelina Rogowska-Wrzesinska, Ole N. Jensen, David R. Gang, Jay J. Thelen and Ian Max Møller, date: 09/05/2014, view: 10469, Q&A: 1

One way to study the function of plant mitochondria is to extract them from plant tissues in an uncontaminated, intact and functional form. The reductionist assumption is that the components present in such a preparation and the in vitro measurable functions or activities reliably reflect the in vivo properties of the organelle ...

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Centrifuge Microscopy to Analyze the Sedimentary Movements of Amyloplasts Authors: Masatsugu Toyota, Norifumi Ikeda, Masao Tasaka and Miyo Terao Morita, date: 09/05/2014, view: 5426, Q&A: 0

A centrifuge microscope (CMS) functionally consists of a centrifuge producing a centrifugal force (hypergravity condition) and a microscope making an enlarged image of an object. This combination of equipment allows live-cell imaging during centrifugation. We have developed a new CMS (NSK Ltd.) to observe movements of the plant organelles such as ...

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Gradient Flotation Centrifugation of Chloroplast Membranes Authors: Venkatasalam Shanmugabalaji and Felix Kessler, date: 09/05/2014, view: 6821, Q&A: 0

Plastoglobules are lipoprotein particles physically attached to thylakoids in chloroplasts (Kessler et al., 1999). They are mainly composed of polar lipid, neutral lipids, and proteins (Vidi et al., 2006). Here we used simple sucrose gradient flotation centrifugation method to purify the plastoglobules from total chloroplast ...

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Axenic Culture of Sclerotinia sclerotiorum and Preparation of Sclerotinia Culture Filtrate Elicitor 1 (SCFE1)-containing Fractions, Triggering Immune Responses in Arabidopsis thaliana Authors: Malou Fraiture and Frédéric Brunner, date: 09/05/2014, view: 6130, Q&A: 0

The necrotrophic white mold fungus Sclerotinia sclerotiorum (S. sclerotiorum) is pathogenic to a broad range of plant species, including the Brassicaceae model plant Arabidopsis thaliana (Boland and Hall, 1994; Bolton et al., 2006). In Arabidopsis thaliana (A. thaliana), the semi-purified ...

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