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Volume:4  Issue:17

September 05, 2014

Biochemistry

Analysis of L- and D-Amino Acids Using UPLC

Authors: Yuta Mutaguchi
Yuta MutaguchiAffiliation: Department of Biotechnology, Akita Prefectural University, Akita City, Japan
Bio-protocol author page: a1644
 and Toshihisa Ohshima
Toshihisa OhshimaAffiliation: Department of Biomedical Engineering, Osaka Institute of Technology, Osaka, Japan
For correspondence: ohshima@bme.oit.ac.jp
Bio-protocol author page: a1645
date: 9/5/2014, 3935 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1231 

[Abstract] 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 ...

Article identifier: bio-protocol.org/e1231

Cell Biology

Dye-uptake Experiment through Connexin Hemichannels

Authors: Andrea Puhar
Andrea PuharAffiliation: Inserm U786 and Institut Pasteur, Unité de Pathogénie Microbienne Moléculaire, Paris, France
For correspondence: andrea.puhar@pasteur.fr
Bio-protocol author page: a1611
 and Philippe J. Sansonetti
Philippe J. SansonettiAffiliation: Microbiologie et Maladies Infectieuses, Collège de France, Paris, France
Bio-protocol author page: a1612
date: 9/5/2014, 3674 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1221 

[Abstract] 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 ...

Article identifier: bio-protocol.org/e1221

Microbiology

Metabolite and Fatty Acid Analysis of Yeast Cells and Culture Supernatants

Authors: Liwei Chen
Liwei ChenAffiliation: School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore, Singapore
Bio-protocol author page: a1609
 and Wei Ning Chen
Wei Ning ChenAffiliation: School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore, Singapore
For correspondence: wnchen@ntu.edu.sg
Bio-protocol author page: a1610
date: 9/5/2014, 5099 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1219 

[Abstract] 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 ...

Article identifier: bio-protocol.org/e1219

Induction of Connexin-hemichannel Opening

Authors: Andrea Puhar
Andrea PuharAffiliation: Inserm U786 and Institut Pasteur, Unité de Pathogénie Microbienne Moléculaire, Paris, France
For correspondence: andrea.puhar@pasteur.fr
Bio-protocol author page: a1611
 and Philippe J. Sansonetti
Philippe J. SansonettiAffiliation: Microbiologie et Maladies Infectieuses, Collège de France, Paris, France
Bio-protocol author page: a1612
date: 9/5/2014, 2419 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1220 

[Abstract] 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 ...

Article identifier: bio-protocol.org/e1220

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Purification of the GfsA-3x FLAG Protein Expressed in Aspergillus nidulans

Authors: Takuji Oka
Takuji OkaAffiliation: Department of Applied Microbial Technology, Sojo University, Kumamoto City, Japan
For correspondence: oka@bio.sojo-u.ac.jp
Bio-protocol author page: a1613
Yukako Katafuchi
Yukako KatafuchiAffiliation: Department of Applied Microbial Technology, Sojo University, Kumamoto City, Japan
Bio-protocol author page: a1614
Kohsai Fukuda
Kohsai FukudaAffiliation: Department of Applied Microbial Technology, Sojo University, Kumamoto City, Japan
Bio-protocol author page: a1615
Keisuke Ekino
Keisuke EkinoAffiliation: Department of Applied Microbial Technology, Sojo University, Kumamoto City, Japan
Bio-protocol author page: a1616
Masatoshi Goto
Masatoshi GotoAffiliation: Department of Bioscience and Biotechnology, Kyushu University, Fukuoka City, Japan
Bio-protocol author page: a1617
 and Yoshiyuki Nomura
Yoshiyuki NomuraAffiliation: Department of Applied Microbial Technology, Sojo University, Kumamoto City, Japan
Bio-protocol author page: a1618
date: 9/5/2014, 2647 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1222 

[Abstract] 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 ...

Article identifier: bio-protocol.org/e1222

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Determination of D-galactofuranose Content of Galactomannoproteins in Aspergillus nidulans

Authors: Takuji Oka
Takuji OkaAffiliation: Department of Applied Microbial Technology, Sojo University, Kumamoto City, Japan
For correspondence: oka@bio.sojo-u.ac.jp
Bio-protocol author page: a1613
Yukako Katafuchi
Yukako KatafuchiAffiliation: Department of Applied Microbial Technology, Sojo University, Kumamoto City, Japan
Bio-protocol author page: a1614
Kohsai Fukuda
Kohsai FukudaAffiliation: Department of Applied Microbial Technology, Sojo University, Kumamoto City, Japan
Bio-protocol author page: a1615
Keisuke Ekino
Keisuke EkinoAffiliation: Department of Applied Microbial Technology, Sojo University, Kumamoto City, Japan
Bio-protocol author page: a1616
Masatoshi Goto
Masatoshi GotoAffiliation: Department of Bioscience and Biotechnology, Kyushu University, Fukuoka City, Japan
Bio-protocol author page: a1617
 and Yoshiyuki Nomura
Yoshiyuki NomuraAffiliation: Department of Applied Microbial Technology, Sojo University, Kumamoto City, Japan
Bio-protocol author page: a1618
date: 9/5/2014, 2136 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1223 

[Abstract] 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 ...

Article identifier: bio-protocol.org/e1223

Iodine Staining of Escherichia coli Expressing Genes Involved in the Synthesis of Bacterial Glycogen

Authors: Ana M. Demonte
Ana M. DemonteAffiliation: Biochemistry and Biological Sciences, National University of the Litoral, Santa Fe, Argentina
Bio-protocol author page: a1619
Matías D. Asención Diez
Matías D. Asención DiezAffiliation: National University of the Litoral, National Council for Research in Science and Technology (CONICET), Santa Fe, Argentina
Bio-protocol author page: a1620
Sergio A. Guerrero
Sergio A. GuerreroAffiliation: Universidad Nacional del Litora, Argentina and Researcher of the National Council of Science (CONICET), Santa Fe, Argentina
Bio-protocol author page: a1621
Miguel A. Ballicora
Miguel A. BallicoraAffiliation: Department of Chemistry and Biochemistry, Loyola University, Chicago, USA
Bio-protocol author page: a1622
 and Alberto A. Iglesias
Alberto A. IglesiasAffiliation: Instituto de Agrobiotecnología del Litoral (UNL-CONICET), Facultad de Bioquímica y Ciencias Biológicas, Santa Fe, Argentina
For correspondence: iglesias@fbcb.unl.edu.ar
Bio-protocol author page: a1623
date: 9/5/2014, 3276 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1224 

[Abstract] 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 ...

Article identifier: bio-protocol.org/e1224

Evolution of Escherichia coli to Macrophage Cell Line

Authors: Migla Miskinyte
Migla MiskinyteAffiliation: Evolutionary Biology Group, Instituto Gulbenkian de Ciência, Oeiras, Portugal
Bio-protocol author page: a1636
 and Isabel Gordo
Isabel GordoAffiliation: Evolutionary Biology Group, Instituto Gulbenkian de Ciência, Oeiras, Portugal
For correspondence: igordo@igc.gulbenkian.pt
Bio-protocol author page: a1637
date: 9/5/2014, 3121 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1227 

[Abstract] 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, ...

Article identifier: bio-protocol.org/e1227

Fitness Measurements of Evolved Esherichia coli

Authors: Migla Miskinyte
Migla MiskinyteAffiliation: Evolutionary Biology Group, Instituto Gulbenkian de Ciência, Oeiras, Portugal
Bio-protocol author page: a1636
 and Isabel Gordo
Isabel GordoAffiliation: Evolutionary Biology Group, Instituto Gulbenkian de Ciência, Oeiras, Portugal
For correspondence: igordo@igc.gulbenkian.pt
Bio-protocol author page: a1637
date: 9/5/2014, 3214 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1228 

[Abstract] 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 ...

Article identifier: bio-protocol.org/e1228

Plant Science

Targeted Gene Mutation in Rice Using a CRISPR-Cas9 System

Authors: Kabin Xie
Kabin XieAffiliation: Department of Plant Pathology and Environmental Microbiology, The Pennsylvania State University, University Park, State College, USA
Bio-protocol author page: a1624
Bastian Minkenberg
Bastian MinkenbergAffiliation: Department of Plant Biology, The Pennsylvania State University, University Park, State College, USA
Bio-protocol author page: a1625
 and Yinong Yang
Yinong YangAffiliation: Department of Plant Biology, The Pennsylvania State University, University Park, State College, USA
For correspondence: yuy3@psu.edu
Bio-protocol author page: a1626
date: 9/5/2014, 10871 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1225 

[Abstract] 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 ...

Article identifier: bio-protocol.org/e1225

Isolation of Mitochondria from Potato Tubers

Authors: Jesper F. Havelund
Jesper F. HavelundAffiliation 1: Department of Molecular Biology and Genetics, Science and Technology, Aarhus University, Slagelse, Denmark
Affiliation 2: Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense M, Denmark
Bio-protocol author page: a1627
Fernanda Salvato
Fernanda SalvatoAffiliation 1: Department of Biochemistry and Interdisciplinary Plant Group, University of Missouri-Columbia, Columbia, USA
Affiliation 2: Institute of Biology, State University of Campinas, Campinas, Brazil
Bio-protocol author page: a1628
Mingjie Chen
Mingjie ChenAffiliation: Department of Biochemistry and Interdisciplinary Plant Group, University of Missouri-Colum, Columbia, USA
Bio-protocol author page: a1629
R.S.P. Rao
R.S.P. RaoAffiliation: Department of Biochemistry and Interdisciplinary Plant Group, University of Missouri-Columbia, Columbia, USA
Bio-protocol author page: a1630
Adelina Rogowska-Wrzesinska
Adelina Rogowska-WrzesinskaAffiliation: Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense M, Denmark
Bio-protocol author page: a1631
Ole N. Jensen
Ole N. JensenAffiliation: Department of Biochemistry and Interdisciplinary Plant Group, University of Southern Denmark, Odense M, Denmark
Bio-protocol author page: a1632
David R. Gang
David R. GangAffiliation: Institute of Biological Chemistry, Washington State University, WA, USA
Bio-protocol author page: a1633
Jay J. Thelen
Jay J. ThelenAffiliation: Department of Biochemistry and Interdisciplinary Plant Group, University of Missouri-Columbia, Columbia, USA
Bio-protocol author page: a1634
 and Ian Max Møller
Ian Max MøllerAffiliation: Department of Molecular Biology and Genetics, Science and Technology, Aarhus University, Slagelse, Denmark
For correspondence: ian.max.moller@agrsci.dk
Bio-protocol author page: a1635
date: 9/5/2014, 5339 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1226 

[Abstract] 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 ...

Article identifier: bio-protocol.org/e1226

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Centrifuge Microscopy to Analyze the Sedimentary Movements of Amyloplasts

Authors: Masatsugu Toyota
Masatsugu ToyotaAffiliation: Department of Botany, University of Wisconsin, Madison, USA
Bio-protocol author page: a1638
Norifumi Ikeda
Norifumi IkedaAffiliation: Mechatronics Technology Development Center, NSK Ltd., Fujisawa, Japan
Bio-protocol author page: a1639
Masao Tasaka
Masao TasakaAffiliation: Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Japan
Bio-protocol author page: a1640
 and Miyo Terao Morita
Miyo Terao MoritaAffiliation: Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan
For correspondence: mimorita@agr.nagoya-u.ac.jp
Bio-protocol author page: a1641
date: 9/5/2014, 2501 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1229 

[Abstract] 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. ...

Article identifier: bio-protocol.org/e1229

Gradient Flotation Centrifugation of Chloroplast Membranes

Authors: Venkatasalam Shanmugabalaji
Venkatasalam ShanmugabalajiAffiliation: Department of Plant Biology, University of Geneva, Geneva, Switzerland
For correspondence: shanmugabalaji.venkatasalam@unige.ch
Bio-protocol author page: a1642
 and Felix Kessler
Felix KesslerAffiliation: Laboratoire de Physiologie Végétale, Université de Neuchâtel, Neuchâtel, Switzerland
Bio-protocol author page: a1643
date: 9/5/2014, 3319 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1230 

[Abstract] 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 ...

Article identifier: bio-protocol.org/e1230

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
Malou FraitureAffiliation: Communication Service, CRP-Santé, Strassen, Luxembourg
Bio-protocol author page: a1646
 and Frédéric Brunner
Frédéric BrunnerAffiliation: Department of Plant Biochemistry, Eberhard Karls University, Tübingen, Germany
For correspondence: frederic.brunner@zmbp.uni-tuebingen.de
Bio-protocol author page: a1647
date: 9/5/2014, 3367 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1232 

[Abstract] 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 ...

Article identifier: bio-protocol.org/e1232