umu.sePublikationer
Ändra sökning
RefereraExporteraLänk till posten
Permanent länk

Direktlänk
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annat format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annat språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
Expression and purification of SfaX(II), a protein involved in regulating adhesion and motility genes in extraintestinal pathogenic Escherichia coli
Umeå universitet, Medicinska fakulteten, Institutionen för molekylärbiologi (Medicinska fakulteten). (UCMR)
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen. (UCMR)
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen. (UCMR)
Umeå universitet, Medicinska fakulteten, Molekylär Infektionsmedicin, Sverige (MIMS). Umeå universitet, Medicinska fakulteten, Institutionen för molekylärbiologi (Medicinska fakulteten). (UCMR)
2012 (Engelska)Ingår i: Protein Expression and Purification, ISSN 1046-5928, E-ISSN 1096-0279, Vol. 86, nr 2, s. 127-134Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Pathogenic Escherichia coli strains commonly harbor genes involved in formation of fimbriae, such as the sfa(II) fimbrial gene cluster found in uropathogenic and newborn meningitis isolates. The sfaX(II) gene, located at the distal end of the sfa(II) operon, was recently shown to play a role in controlling virulence-related gene expression in extraintestinal pathogenic E. coli (ExPEC). Until now, detailed characterization of the SfaX(II) protein has been hampered by difficulties in obtaining large quantities of soluble protein. By a rational modeling approach, we engineered a Cys70Ser mutation, which successfully improved solubility of the protein. Here, we present the expression, purification, and initial characterization of the recombinant SfaX(IIC70S) mutant. The protein was produced in E. coli BL21 (DE3) cells grown in autoinduction culture media. The plasmid vector harbored DNA encoding the SfaX(IIC70S) protein N-terminally fused with a six histidine (H6) sequence followed by a ZZ tag (a derivative of the Staphylococcus protein A) (H6-ZZ tag). The H6-ZZ tag was cleaved off with Tobacco Etch Virus (TEV) protease and the 166 amino acid full-length homo-dimeric protein was purified using affinity and size-exclusion chromatography. Electrophoretic mobility gel shift assays and atomic force microscopy demonstrated that the protein possesses DNA-binding properties, suggesting that the transcriptional regulatory activity of SfaX(II) can be mediated via direct binding to DNA.

Ort, förlag, år, upplaga, sidor
Elsevier, 2012. Vol. 86, nr 2, s. 127-134
Nyckelord [en]
SfaXII, ExPEC, Fimbriae, Transcription regulation
Nationell ämneskategori
Kemi Medicin och hälsovetenskap
Identifikatorer
URN: urn:nbn:se:umu:diva-60770DOI: 10.1016/j.pep.2012.09.006PubMedID: 23022032OAI: oai:DiVA.org:umu-60770DiVA, id: diva2:562794
Tillgänglig från: 2012-10-26 Skapad: 2012-10-26 Senast uppdaterad: 2018-06-08Bibliografiskt granskad

Open Access i DiVA

Fulltext saknas i DiVA

Övriga länkar

Förlagets fulltextPubMed

Personposter BETA

Paracuellos, PatriciaÖhman, AndersSauer-Eriksson, A ElisabethUhlin, Bernt Eric

Sök vidare i DiVA

Av författaren/redaktören
Paracuellos, PatriciaÖhman, AndersSauer-Eriksson, A ElisabethUhlin, Bernt Eric
Av organisationen
Institutionen för molekylärbiologi (Medicinska fakulteten)Kemiska institutionenMolekylär Infektionsmedicin, Sverige (MIMS)
I samma tidskrift
Protein Expression and Purification
KemiMedicin och hälsovetenskap

Sök vidare utanför DiVA

GoogleGoogle Scholar

doi
pubmed
urn-nbn

Altmetricpoäng

doi
pubmed
urn-nbn
Totalt: 336 träffar
RefereraExporteraLänk till posten
Permanent länk

Direktlänk
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annat format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annat språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf