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Article: An optimized protocol for isolation of soluble proteins from microalgae for two-dimensional gel electrophoresis analysis

TitleAn optimized protocol for isolation of soluble proteins from microalgae for two-dimensional gel electrophoresis analysis
Authors
KeywordsGreen Algae
Haematococcus Pluvialis
Proteomics
Thiourea
Tributyl Phosphine
Two-Dimensional Gel Electrophoresis
Issue Date2003
PublisherSpringer Verlag Dordrecht. The Journal's web site is located at http://springerlink.metapress.com/openurl.asp?genre=journal&issn=0921-8971
Citation
Journal Of Applied Phycology, 2003, v. 15 n. 6, p. 485-496 How to Cite?
AbstractTwo-dimensional gel electrophoresis (2-DE) is a core proteomic technique to study protein expression and function in living organisms. Although it has been extensively used for investigation of bacterial, yeast, animal and plant tissue cells, there is limited information about the use of 2-DE in microalgal research. In this study, a number of key chemical reagents, including acetone, trichloroacetic acid, urea, thiourea, dithiothreitol, and tributyl phosphine, were quantitatively evaluated for 2-DE of green microalgae, using Haematococcus pluvialis as a model system. The goal was to maximize the number and staining intensity of protein spots while minimizing streaking and smearing on the second dimensional SDS gel. Compared to non-frozen immobilized pH gradients (IPG) strips, freezing of the IPG strips at -20°C after isoelectric focusing (IEF) enhanced protein resolubilization and transfer into the SDS gel, and thus improved resolution while eliminating vertical point streaking on the SDS gel. It was also confirmed that manipulation of sample loading capacity is a simple, effective purification strategy for selective investigation of the proteins of interest and of varying abundances. The protocol was also successfully applied to profiling protein expression in H. pluvialis under external stress conditions, indicating its potential usefulness in further proteomics studies of this organism and related species.
Persistent Identifierhttp://hdl.handle.net/10722/178841
ISSN
2015 Impact Factor: 2.372
2015 SCImago Journal Rankings: 0.852
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorWang, SBen_US
dc.contributor.authorHu, Qen_US
dc.contributor.authorSommerfeld, Men_US
dc.contributor.authorChen, Fen_US
dc.date.accessioned2012-12-19T09:50:04Z-
dc.date.available2012-12-19T09:50:04Z-
dc.date.issued2003en_US
dc.identifier.citationJournal Of Applied Phycology, 2003, v. 15 n. 6, p. 485-496en_US
dc.identifier.issn0921-8971en_US
dc.identifier.urihttp://hdl.handle.net/10722/178841-
dc.description.abstractTwo-dimensional gel electrophoresis (2-DE) is a core proteomic technique to study protein expression and function in living organisms. Although it has been extensively used for investigation of bacterial, yeast, animal and plant tissue cells, there is limited information about the use of 2-DE in microalgal research. In this study, a number of key chemical reagents, including acetone, trichloroacetic acid, urea, thiourea, dithiothreitol, and tributyl phosphine, were quantitatively evaluated for 2-DE of green microalgae, using Haematococcus pluvialis as a model system. The goal was to maximize the number and staining intensity of protein spots while minimizing streaking and smearing on the second dimensional SDS gel. Compared to non-frozen immobilized pH gradients (IPG) strips, freezing of the IPG strips at -20°C after isoelectric focusing (IEF) enhanced protein resolubilization and transfer into the SDS gel, and thus improved resolution while eliminating vertical point streaking on the SDS gel. It was also confirmed that manipulation of sample loading capacity is a simple, effective purification strategy for selective investigation of the proteins of interest and of varying abundances. The protocol was also successfully applied to profiling protein expression in H. pluvialis under external stress conditions, indicating its potential usefulness in further proteomics studies of this organism and related species.en_US
dc.languageengen_US
dc.publisherSpringer Verlag Dordrecht. The Journal's web site is located at http://springerlink.metapress.com/openurl.asp?genre=journal&issn=0921-8971en_US
dc.relation.ispartofJournal of Applied Phycologyen_US
dc.subjectGreen Algaeen_US
dc.subjectHaematococcus Pluvialisen_US
dc.subjectProteomicsen_US
dc.subjectThioureaen_US
dc.subjectTributyl Phosphineen_US
dc.subjectTwo-Dimensional Gel Electrophoresisen_US
dc.titleAn optimized protocol for isolation of soluble proteins from microalgae for two-dimensional gel electrophoresis analysisen_US
dc.typeArticleen_US
dc.identifier.emailChen, F: sfchen@hku.hken_US
dc.identifier.authorityChen, F=rp00672en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1023/B:JAPH.0000004324.88897.b2en_US
dc.identifier.scopuseid_2-s2.0-0348011459en_US
dc.identifier.hkuros90932-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0348011459&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume15en_US
dc.identifier.issue6en_US
dc.identifier.spage485en_US
dc.identifier.epage496en_US
dc.identifier.isiWOS:000186637300005-
dc.publisher.placeNetherlandsen_US
dc.identifier.scopusauthoridWang, SB=7410347419en_US
dc.identifier.scopusauthoridHu, Q=26666082400en_US
dc.identifier.scopusauthoridSommerfeld, M=7007025132en_US
dc.identifier.scopusauthoridChen, F=7404907980en_US

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