2.50
Hdl Handle:
http://hdl.handle.net/10033/84758
Title:
Heterologous expression of the msp2 gene from Marasmius scorodonius.
Authors:
Zelena, Kateryna; Zorn, Holger; Nimtz, Manfred; Berger, Ralf Günter
Abstract:
For the heterologous expression of the msp2 gene from the edible mushroom Marasmius scorodonius in Escherichia coli the cDNA encoding the extracellular Msp2 peroxidase was cloned into the pBAD III expression plasmid. Expression of the protein with or without signal peptide was investigated in E. coli strains TOP10 and LMG194. Different PCR products were amplified for expression of the native target protein or a protein with a signal peptide. Omitting the native stop codon and adding six His-residues resulted in a fusion protein amenable to immune detection and purification by immobilised metal affinity chromatography. In E. coli the recombinant protein was produced in high yield as insoluble inclusion bodies. The influence of different parameters on MsP2 refolding was investigated. Active enzyme was obtained by glutathione-mediated oxidation in a medium containing urea, Ca(2+), and hemin.
Affiliation:
Institut für Lebensmittelchemie, Leibniz Universität Hannover, Callinstrasse 5, Hannover, Germany.
Citation:
Heterologous expression of the msp2 gene from Marasmius scorodonius. 2009, 191 (5):397-402 Arch. Microbiol.
Journal:
Archives of microbiology
Issue Date:
May-2009
URI:
http://hdl.handle.net/10033/84758
DOI:
10.1007/s00203-009-0462-2
PubMed ID:
19247632
Type:
Article
Language:
en
ISSN:
1432-072X
Appears in Collections:
publications of the research group cellular proteom research (CPRO)

Full metadata record

DC FieldValue Language
dc.contributor.authorZelena, Katerynaen
dc.contributor.authorZorn, Holgeren
dc.contributor.authorNimtz, Manfreden
dc.contributor.authorBerger, Ralf Günteren
dc.date.accessioned2009-10-23T11:40:16Z-
dc.date.available2009-10-23T11:40:16Z-
dc.date.issued2009-05-
dc.identifier.citationHeterologous expression of the msp2 gene from Marasmius scorodonius. 2009, 191 (5):397-402 Arch. Microbiol.en
dc.identifier.issn1432-072X-
dc.identifier.pmid19247632-
dc.identifier.doi10.1007/s00203-009-0462-2-
dc.identifier.urihttp://hdl.handle.net/10033/84758-
dc.description.abstractFor the heterologous expression of the msp2 gene from the edible mushroom Marasmius scorodonius in Escherichia coli the cDNA encoding the extracellular Msp2 peroxidase was cloned into the pBAD III expression plasmid. Expression of the protein with or without signal peptide was investigated in E. coli strains TOP10 and LMG194. Different PCR products were amplified for expression of the native target protein or a protein with a signal peptide. Omitting the native stop codon and adding six His-residues resulted in a fusion protein amenable to immune detection and purification by immobilised metal affinity chromatography. In E. coli the recombinant protein was produced in high yield as insoluble inclusion bodies. The influence of different parameters on MsP2 refolding was investigated. Active enzyme was obtained by glutathione-mediated oxidation in a medium containing urea, Ca(2+), and hemin.en
dc.language.isoenen
dc.subject.meshCalciumen
dc.subject.meshChromatography, Affinityen
dc.subject.meshCloning, Molecularen
dc.subject.meshCoenzymesen
dc.subject.meshDNA, Complementaryen
dc.subject.meshEnzyme Stabilityen
dc.subject.meshEscherichia colien
dc.subject.meshFungal Proteinsen
dc.subject.meshGene Expressionen
dc.subject.meshGenetic Vectorsen
dc.subject.meshGlutathioneen
dc.subject.meshHeminen
dc.subject.meshHydrogen-Ion Concentrationen
dc.subject.meshMarasmiusen
dc.subject.meshOxidantsen
dc.subject.meshOxidation-Reductionen
dc.subject.meshPeroxidaseen
dc.subject.meshProtein Sorting Signalsen
dc.subject.meshRecombinant Fusion Proteinsen
dc.titleHeterologous expression of the msp2 gene from Marasmius scorodonius.en
dc.typeArticleen
dc.contributor.departmentInstitut für Lebensmittelchemie, Leibniz Universität Hannover, Callinstrasse 5, Hannover, Germany.en
dc.identifier.journalArchives of microbiologyen

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