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High-level expression of Bacillus stearothermophilus 6-phosphofructo-1-kinase in Escherichia coli
B A French1, B C Valdez, E S Younathan
1Department of Biochemistry, Louisiana State University, Baton Rouge 70803.
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.