title: High-level expression of Bacillus stearothermophilus 6-phosphofructo-1-kinase in Escherichia coli.
abstract: The 6-phosphofructo-1-kinase (PFK) gene from Bacillus stearothermophilus has been expressed at high levels in Escherichia coli. This expression has been demonstrated by complementation studies, sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, and PFK assays of cell extracts. A level of B. stearothermophilus PFK expression corresponding to 20% of the total extracted protein was calculated from densitometric scans of an SDS-polyacrylamide gel. The high level of recombinant gene expression will enable this laboratory to determine structure-function relationships in B. stearothermophilus PFK by the method of oligodeoxynucleotide-directed mutagenesis.
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D001411:Geobacillus stearothermophilus / Q000201:enzymology / Q000235:genetics*; D010732:Phosphofructokinase-1 / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D010732:Phosphofructokinase-1