|
rdf:type |
|
|
lifeskim:mentions |
|
|
pubmed:issue |
5
|
|
pubmed:dateCreated |
1991-10-30
|
|
pubmed:abstractText |
We have constructed a shuttle plasmid for Bacillus megaterium and Escherichia coli that contains the promoter and repressor gene of the B. megaterium-borne operon for xylose utilization. A polylinker downstream of the promoter allows versatile cloning of genes under its transcriptional control. We have placed gdhA (encoding glucose dehydrogenase) from B. megaterium, lacZ (encoding beta-galactosidase) from E. coli, mro (encoding mutarotase) from Acinetobacter calcoaceticus, and human puk (encoding single-chain urokinase-like plasminogen activator, rscuPA) under xylose control in this vector. All four genes were between 130-fold and 350-fold inducible by 0.5% xylose in the growth medium in B. megaterium. Enzymatically active glucose dehydrogenase and mutarotase accumulated to 20% and 30% of the total soluble protein, respectively. beta-Galactosidase and rscuPA were also expressed at a high level. A gel analysis of the products demonstrated their proteolytic stability in the cytoplasm, even up to 5 h after induction. The expression properties of this new host-vector system are discussed in comparison to the ones available for B. subtilis and E. coli.
|
|
pubmed:language |
eng
|
|
pubmed:journal |
|
|
pubmed:citationSubset |
B
|
|
pubmed:chemical |
|
|
pubmed:status |
MEDLINE
|
|
pubmed:month |
Aug
|
|
pubmed:issn |
0175-7598
|
|
pubmed:author |
|
|
pubmed:issnType |
Print
|
|
pubmed:volume |
35
|
|
pubmed:geneSymbol |
gdhA,
lacZ,
mro,
puk
|
|
pubmed:owner |
NLM
|
|
pubmed:authorsComplete |
Y
|
|
pubmed:pagination |
594-9
|
|
pubmed:dateRevised |
2008-11-21
|
|
pubmed:meshHeading |
pubmed-meshheading:1367576-Acinetobacter,
pubmed-meshheading:1367576-Amino Acid Sequence,
pubmed-meshheading:1367576-Bacillus megaterium,
pubmed-meshheading:1367576-Base Sequence,
pubmed-meshheading:1367576-Carbohydrate Epimerases,
pubmed-meshheading:1367576-Cloning, Molecular,
pubmed-meshheading:1367576-Enzyme Precursors,
pubmed-meshheading:1367576-Escherichia coli,
pubmed-meshheading:1367576-Glucose 1-Dehydrogenase,
pubmed-meshheading:1367576-Glucose Dehydrogenases,
pubmed-meshheading:1367576-Humans,
pubmed-meshheading:1367576-Molecular Sequence Data,
pubmed-meshheading:1367576-Operon,
pubmed-meshheading:1367576-Plasmids,
pubmed-meshheading:1367576-Recombinant Fusion Proteins,
pubmed-meshheading:1367576-Regulatory Sequences, Nucleic Acid,
pubmed-meshheading:1367576-Urokinase-Type Plasminogen Activator,
pubmed-meshheading:1367576-Xylose,
pubmed-meshheading:1367576-beta-Galactosidase
|
|
pubmed:year |
1991
|
|
pubmed:articleTitle |
Inducible high-level expression of heterologous genes in Bacillus megaterium using the regulatory elements of the xylose-utilization operon.
|
|
pubmed:affiliation |
Lehrstuhl für Mikrobiologie, Institut für Mikrobiologie und Biochemieder Friedrich-Alexander Universität Erlangen-Nürnberg, Federal Republic of Germany.
|
|
pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|
