Identification, isolation, and overexpression of the gene encoding the psi subunit of DNA polymerase III holoenzyme.
The gene encoding the psi subunit of DNA polymerase III holoenzyme, holD, was identified and isolated by an approach in which peptide sequence data were used to obtain a DNA hybridization probe. The gene, which maps to 99.3 centisomes, was sequenced and found to be identical to a previously uncharacterized open reading frame that overlaps the 5' end of rimI by 29 bases, contains 411 bp, and is predicted to encode a protein of 15,174 Da. When expressed in a plasmid that also expressed holC, holD directed expression of the psi subunit to about 3% of total soluble protein.
Carter
J R
JR
Department of Biochemistry, Biophysics and Genetics, University of Colorado Health Sciences Center, Denver 80262.
Franden
M A
MA
Aebersold
R
R
McHenry
C S
CS
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004258:DNA Polymerase III / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010957:Plasmids; D016133:Polymerase Chain Reaction
D004269:DNA, Bacterial; D004258:DNA Polymerase III