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Rubisco Extraction and Purification from Diatoms
Authors:
Jodi N. Young
Jodi N. Young
Affiliation:
Department of Oceanography, University of Washington, Seattle, USA
For correspondence:
youngjn@uw.edu
Bio-protocol author page:
a4026
,
Ana M. C. Heureux
Ana M. C. Heureux
Affiliation:
Department of Earth Sciences, University of Oxford, Oxford, UK
Bio-protocol author page:
a4027
,
Rosalind E. M. Rickaby
Rosalind E. M. Rickaby
Affiliation:
Department of Earth Sciences, University of Oxford, Oxford, UK
Bio-protocol author page:
a4028
,
François M. M. Morel
François M. M. Morel
Affiliation:
Department of Geosciences, Princeton University, Princeton, USA
Bio-protocol author page:
a4029
,
Spencer M. Whitney
Spencer M. Whitney
Affiliation:
Plant Science Division, Research School of Biology, the Australian National University, Canberra, Australia
Bio-protocol author page:
a4030
and
Robert E. Sharwood
Robert E. Sharwood
Affiliation:
Plant Science Division, Research School of Biology, the Australian National University, Canberra, Australia
Bio-protocol author page:
a4031
,
date: 3/20/2017, 668 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.2191
.
[Abstract] This protocol describes a method to extract ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco) from diatoms (
Bacillariophyta
) to determine catalytic performance. This protocol has been adapted from use in cyanobacteria and higher plants (Andrews, 1988; Whitney and Sharwood, 2007). First part (steps A1-A3) of the extraction provides a crude extract ...
Chlorophyll Fluorescence Measurements in
Arabidopsis
Plants Using a Pulse-amplitude-modulated (PAM) Fluorometer
Authors:
Reiko Motohashi
Reiko Motohashi
Affiliation:
Agriculture Department, Shizuoka University, Shizuoka, Japan
For correspondence:
armotoh@ipc.shizuoka.ac.jp
Bio-protocol author page:
a2179
and
Fumiyoshi Myouga
Fumiyoshi Myouga
Affiliation:
RIKEN Center for Sustainable Resource Science, Yokohama, Japan
Bio-protocol author page:
a2181
,
date: 5/5/2015, 4627 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1464
.
[Abstract] In this protocol, to analyze PSII activity in photosynthesis, we measure the Fv/Fm (Fv=Fm ± Fo) value (Fo and Fm are the minimum and maximum values of chlorophyll fluorescence of dark-adapted leaves, respectively). Fv/Fm is a reliable marker of photo- inhibition (Krause
et al.
, 1988). Chlorophyll fluorescence in leaves was measured at room temperature ...
Measurement of Chlorophyll a and Carotenoids Concentration in Cyanobacteria
Authors:
Tomáš Zavřel
Tomáš Zavřel
Affiliation:
Department of Adaptation Biotechnologies, Global Change Research Centre, Academy of Science of the Czech Republic, Drásov, Czech Republic
For correspondence:
zavrel.t@czechglobe.cz
Bio-protocol author page:
a2192
,
Maria A. Sinetova
Maria A. Sinetova
Affiliation:
Laboratory of Intracellular Regulation, Institute of Plant Physiology, Russian Academy of Sciences, Moscow, Russia
Bio-protocol author page:
a243
and
Jan Červený
Jan Červený
Affiliation:
Department of Adaptation Biotechnologies, Global Change Research Centre, Academy of Science of the Czech Republic, Drásov, Czech Republic
Bio-protocol author page:
a2193
,
date: 5/5/2015, 6599 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1467
.
[Abstract] This is a protocol for precise measurement of chlorophyll
a
and total carotenoid concentrations in cyanobacteria cells. Cellular chlorophyll concentration is one of the central physiological parameters, routinely followed in many research areas ranging from stress physiology to biotechnology. Carotenoids concentration is often related to cellular stress ...
Detection and Quantification of Heme and Chlorophyll Precursors Using a High Performance Liquid Chromatography (HPLC) System Equipped with Two Fluorescence Detectors
Authors:
Jan Pilný
Jan Pilný
Affiliation:
Institute of Microbiology, Academy of Sciences, Třeboň, Czech Republic
Bio-protocol author page:
a1970
,
Jana Kopečná
Jana Kopečná
Affiliation:
Institute of Microbiology, Academy of Sciences, Třeboň, Czech Republic
Bio-protocol author page:
a86
,
Judith Noda
Judith Noda
Affiliation:
Institute of Microbiology, Academy of Sciences, Třeboň, Czech Republic
Bio-protocol author page:
a1971
and
Roman Sobotka
Roman Sobotka
Affiliation:
Institute of Microbiology, Academy of Sciences, Třeboň, Czech Republic
For correspondence:
sobotka@alga.cz
Bio-protocol author page:
a84
,
date: 2/5/2015, 3252 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1390
.
[Abstract] Intermediates of tetrapyrrole biosynthetic pathway are low-abundant compounds, and their quantification is usually difficult, time consuming and requires large amounts of input material. Here, we describe a method allowing fast and accurate quantification of almost all intermediates of the heme and chlorophyll biosynthesis, including mono-vinyl and ...
Analysis of Malondialdehyde, Chlorophyll Proline, Soluble Sugar, and Glutathione Content in
Arabidopsis
seedling
Authors:
Zhijin Zhang
Zhijin Zhang
Affiliation:
Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou, China
Bio-protocol author page:
a684
and
Rongfeng Huang
Rongfeng Huang
Affiliation:
Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou, China
For correspondence:
rfhuang@caas.cn
Bio-protocol author page:
a362
,
date: 7/20/2013, 11118 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.817
.
[Abstract] The protocol has four sub-protocols, which are about the measurement of malondialdehyde, chlorophyll proline, soluble sugar, and glutathione content, respectively, in
Arabidopsis
seedling by using spectrophotometer. These methods are simple, effective and reproducible, which will help the researchers who are not familiar with these approaches, quickly ...
35
S pulse Labelling of
Chlamydomonas
Chloroplast Proteins
Authors:
Alexandra-Viola Bohne
Alexandra-Viola Bohne
Affiliation:
Molecular Plant Science, Ludwig-Maximilians-Universität, Munich, Germany
Bio-protocol author page:
a615
,
Christian Schwarz
Christian Schwarz
Affiliation:
Molecular Plant Science, Ludwig-Maximilians-Universität, Munich, Germany
Bio-protocol author page:
a609
and
Joerg Nickelsen
Joerg Nickelsen
Affiliation:
Molecular Plant Science, Ludwig-Maximilians-Universität, Munich, Germany
For correspondence:
joerg.nickelsen@lrz.uni-muenchen.de
Bio-protocol author page:
a610
,
date: 6/5/2013, 3952 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.783
.
[Abstract]
35
S pulse labelling of proteins is used to attach a radioactive label to newly synthesized proteins, as sulfur is an element that is mainly present in proteins (Fleischmann and Rochaix 1999). Depending on your organism’s uptake mechanisms you need cysteine, methionine or sulfuric acid as a source of radioactive sulfur. This example uses
Chlamydomonas
...
Radiolabeling of Chlorophyll by [14C]Glutamic Acid
in vivo
and Relative Quantification of Labeled Chlorophyll by Using Thin Layer Chromatography (TLC)
Authors:
Roman Sobotka
Roman Sobotka
Affiliation:
Department of Phototrophic Microorganisms, Institute of Microbiology ASCR, Trebon, Czech Republic
For correspondence:
sobotka@alga.cz
Bio-protocol author page:
a84
,
Luděk Kořený
Luděk Kořený
Affiliation:
Department of Molecular Parasitology, Institute of Parasitology, Biology Centre ASCR, Ceske Budejovice, Czech Republic
Bio-protocol author page:
a85
,
Jana Kopečná
Jana Kopečná
Affiliation:
Department of Phototrophic Microorganisms, Institute of Microbiology ASCR, Trebon, Czech Republic
Bio-protocol author page:
a86
and
Miroslav Oborník
Miroslav Oborník
Affiliation:
Department of Molecular Parasitology, Institute of Parasitology, Biology Centre ASCR, Ceske Budejovice, Czech Republic
Bio-protocol author page:
a87
,
date: 8/20/2012, 3989 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.249
.
[Abstract] This is an accurate method to assess the rate of chlorophyll biosynthesis
in vivo
in cyanobacteria. Given that labeled glutamate is used as the very early precursor of chlorophyll together with a short pulse of labeling (30 min), this method provides information about the metabolic flow through the whole chlorophyll biosynthetic pathway on a short ...
1
Analysis of Malondialdehyde, Chlorophyll Proline, Soluble Sugar, and Glutathione Content in
Arabidopsis
seedling
Authors:
Zhijin Zhang
Zhijin Zhang
Affiliation:
Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou, China
Bio-protocol author page:
a684
and
Rongfeng Huang
Rongfeng Huang
Affiliation:
Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou, China
For correspondence:
rfhuang@caas.cn
Bio-protocol author page:
a362
,
date: 7/20/2013, 11118 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.817
.
[Abstract] The protocol has four sub-protocols, which are about the measurement of malondialdehyde, chlorophyll proline, soluble sugar, and glutathione content, respectively, in
Arabidopsis
seedling by using spectrophotometer. These methods are simple, effective and reproducible, which will help the researchers ...
Measurement of Chlorophyll a and Carotenoids Concentration in Cyanobacteria
Authors:
Tomáš Zavřel
Tomáš Zavřel
Affiliation:
Department of Adaptation Biotechnologies, Global Change Research Centre, Academy of Science of the Czech Republic, Drásov, Czech Republic
For correspondence:
zavrel.t@czechglobe.cz
Bio-protocol author page:
a2192
,
Maria A. Sinetova
Maria A. Sinetova
Affiliation:
Laboratory of Intracellular Regulation, Institute of Plant Physiology, Russian Academy of Sciences, Moscow, Russia
Bio-protocol author page:
a243
and
Jan Červený
Jan Červený
Affiliation:
Department of Adaptation Biotechnologies, Global Change Research Centre, Academy of Science of the Czech Republic, Drásov, Czech Republic
Bio-protocol author page:
a2193
,
date: 5/5/2015, 6599 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1467
.
[Abstract] This is a protocol for precise measurement of chlorophyll
a
and total carotenoid concentrations in cyanobacteria cells. Cellular chlorophyll concentration is one of the central physiological parameters, routinely followed in many research areas ranging from stress physiology to biotechnology. Carotenoids ...
Chlorophyll Fluorescence Measurements in
Arabidopsis
Plants Using a Pulse-amplitude-modulated (PAM) Fluorometer
Authors:
Reiko Motohashi
Reiko Motohashi
Affiliation:
Agriculture Department, Shizuoka University, Shizuoka, Japan
For correspondence:
armotoh@ipc.shizuoka.ac.jp
Bio-protocol author page:
a2179
and
Fumiyoshi Myouga
Fumiyoshi Myouga
Affiliation:
RIKEN Center for Sustainable Resource Science, Yokohama, Japan
Bio-protocol author page:
a2181
,
date: 5/5/2015, 4627 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1464
.
[Abstract] In this protocol, to analyze PSII activity in photosynthesis, we measure the Fv/Fm (Fv=Fm ± Fo) value (Fo and Fm are the minimum and maximum values of chlorophyll fluorescence of dark-adapted leaves, respectively). Fv/Fm is a reliable marker of photo- inhibition (Krause
et al.
, 1988). Chlorophyll fluorescence ...
Radiolabeling of Chlorophyll by [14C]Glutamic Acid
in vivo
and Relative Quantification of Labeled Chlorophyll by Using Thin Layer Chromatography (TLC)
Authors:
Roman Sobotka
Roman Sobotka
Affiliation:
Department of Phototrophic Microorganisms, Institute of Microbiology ASCR, Trebon, Czech Republic
For correspondence:
sobotka@alga.cz
Bio-protocol author page:
a84
,
Luděk Kořený
Luděk Kořený
Affiliation:
Department of Molecular Parasitology, Institute of Parasitology, Biology Centre ASCR, Ceske Budejovice, Czech Republic
Bio-protocol author page:
a85
,
Jana Kopečná
Jana Kopečná
Affiliation:
Department of Phototrophic Microorganisms, Institute of Microbiology ASCR, Trebon, Czech Republic
Bio-protocol author page:
a86
and
Miroslav Oborník
Miroslav Oborník
Affiliation:
Department of Molecular Parasitology, Institute of Parasitology, Biology Centre ASCR, Ceske Budejovice, Czech Republic
Bio-protocol author page:
a87
,
date: 8/20/2012, 3989 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.249
.
[Abstract] This is an accurate method to assess the rate of chlorophyll biosynthesis
in vivo
in cyanobacteria. Given that labeled glutamate is used as the very early precursor of chlorophyll together with a short pulse of labeling (30 min), this method provides information about the metabolic flow through the ...
35
S pulse Labelling of
Chlamydomonas
Chloroplast Proteins
Authors:
Alexandra-Viola Bohne
Alexandra-Viola Bohne
Affiliation:
Molecular Plant Science, Ludwig-Maximilians-Universität, Munich, Germany
Bio-protocol author page:
a615
,
Christian Schwarz
Christian Schwarz
Affiliation:
Molecular Plant Science, Ludwig-Maximilians-Universität, Munich, Germany
Bio-protocol author page:
a609
and
Joerg Nickelsen
Joerg Nickelsen
Affiliation:
Molecular Plant Science, Ludwig-Maximilians-Universität, Munich, Germany
For correspondence:
joerg.nickelsen@lrz.uni-muenchen.de
Bio-protocol author page:
a610
,
date: 6/5/2013, 3952 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.783
.
[Abstract]
35
S pulse labelling of proteins is used to attach a radioactive label to newly synthesized proteins, as sulfur is an element that is mainly present in proteins (Fleischmann and Rochaix 1999). Depending on your organism’s uptake mechanisms you need cysteine, methionine or sulfuric acid as a source of ...
Detection and Quantification of Heme and Chlorophyll Precursors Using a High Performance Liquid Chromatography (HPLC) System Equipped with Two Fluorescence Detectors
Authors:
Jan Pilný
Jan Pilný
Affiliation:
Institute of Microbiology, Academy of Sciences, Třeboň, Czech Republic
Bio-protocol author page:
a1970
,
Jana Kopečná
Jana Kopečná
Affiliation:
Institute of Microbiology, Academy of Sciences, Třeboň, Czech Republic
Bio-protocol author page:
a86
,
Judith Noda
Judith Noda
Affiliation:
Institute of Microbiology, Academy of Sciences, Třeboň, Czech Republic
Bio-protocol author page:
a1971
and
Roman Sobotka
Roman Sobotka
Affiliation:
Institute of Microbiology, Academy of Sciences, Třeboň, Czech Republic
For correspondence:
sobotka@alga.cz
Bio-protocol author page:
a84
,
date: 2/5/2015, 3252 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1390
.
[Abstract] Intermediates of tetrapyrrole biosynthetic pathway are low-abundant compounds, and their quantification is usually difficult, time consuming and requires large amounts of input material. Here, we describe a method allowing fast and accurate quantification of almost all intermediates of the heme and ...
Rubisco Extraction and Purification from Diatoms
Authors:
Jodi N. Young
Jodi N. Young
Affiliation:
Department of Oceanography, University of Washington, Seattle, USA
For correspondence:
youngjn@uw.edu
Bio-protocol author page:
a4026
,
Ana M. C. Heureux
Ana M. C. Heureux
Affiliation:
Department of Earth Sciences, University of Oxford, Oxford, UK
Bio-protocol author page:
a4027
,
Rosalind E. M. Rickaby
Rosalind E. M. Rickaby
Affiliation:
Department of Earth Sciences, University of Oxford, Oxford, UK
Bio-protocol author page:
a4028
,
François M. M. Morel
François M. M. Morel
Affiliation:
Department of Geosciences, Princeton University, Princeton, USA
Bio-protocol author page:
a4029
,
Spencer M. Whitney
Spencer M. Whitney
Affiliation:
Plant Science Division, Research School of Biology, the Australian National University, Canberra, Australia
Bio-protocol author page:
a4030
and
Robert E. Sharwood
Robert E. Sharwood
Affiliation:
Plant Science Division, Research School of Biology, the Australian National University, Canberra, Australia
Bio-protocol author page:
a4031
,
date: 3/20/2017, 668 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.2191
.
[Abstract] This protocol describes a method to extract ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco) from diatoms (
Bacillariophyta
) to determine catalytic performance. This protocol has been adapted from use in cyanobacteria and higher plants (Andrews, 1988; Whitney and Sharwood, 2007). First part ...
1
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