Linked Life Data

7756834

Source:http://linkedlifedata.com/resource/pubmed/id/7756834

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1995-6-29
pubmed:abstractText
The malK gene of Salmonella typhimurium encoding the ATP-hydrolyzing subunit of the ATP-Binding Cassette (ABC) transporter for maltose was subcloned into the pRSET5d expression vector. Subsequently, the resulting plasmid (pES67) was introduced into Escherichia coli strain BL21(DE3)/pLysS. When strain BL21-(DE3)/pLysS/pES67 was grown at 30 degrees C in a tryptone-phosphate medium (J.T. Moore, A. Uppal, F. Maley, and G. F. Maley, Protein Expression Purif. 4, 160-163, 1993), the addition of isopropyl beta-thiogalactoside resulted in the synthesis of large amounts of MalK protein. After cell disrupture about 60% of MalK was recovered with the soluble (cytoplasmic) fraction. The protein was purified by ion exchange chromatography and dye ligand affinity chromatography. The purified MalK protein displayed enzymatic properties similar to those of a preparation purified and renatured from inclusion bodies (S. Morbach, S. Tebbe, and E. Schneider, J. Biol. Chem. 268, 18617-18621, 1993). Thus, our results disprove the view that the biochemical properties of a protein renatured from inclusion bodies might be artefactual. In addition, we provide further evidence that the modification of growth conditions and the use of a T7 expression system can be a useful approach to overcome at least in part the formation of inclusion bodies.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
1046-5928
pubmed:author
pubmed:issnType
Print
pubmed:volume
6
pubmed:geneSymbol
malK
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
10-4
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
1995
pubmed:articleTitle
Functional purification of a bacterial ATP-binding cassette transporter protein (MalK) from the cytoplasmic fraction of an overproducing strain.
pubmed:affiliation
Arbeitsgruppe Mikrobiologie, Fachbereich Biologie/Chemie, Universität Osnabrück, Germany.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, Non-U.S. Gov't