title: Isolation, sequencing and overexpression of the gene encoding the theta subunit of DNA polymerase III holoenzyme.
abstract: The gene encoding the theta subunit of DNA polymerase III holoenzyme, designated holE, was isolated using a strategy in which peptide sequence was used to derive a DNA hybridization probe. Sequencing of the gene, which maps to 41.43 centisomes of the chromosome, revealed a 76-codon open reading frame predicted to produce a protein of 8,846 Da. When placed in a tac promoter expression vector, the open reading frame directed expression of a protein, that comigrated with authentic theta subunit from purified holoenzyme, to 6% of total soluble protein.
authors.affiliation: University of Colorado Health Sciences Center, Department of Biochemistry, Biophysics and Genetics, Denver 80262.
authors.forename: J R
authors.initials: JR
authors.lastname: Carter
authors.forename: M A
authors.initials: MA
authors.lastname: Franden
authors.forename: R
authors.initials: R
authors.lastname: Aebersold
authors.forename: D R
authors.initials: DR
authors.lastname: Kim
authors.forename: C S
authors.initials: CS
authors.lastname: McHenry
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D004258:DNA Polymerase III / Q000235:genetics* / Q000378:metabolism; D015342:DNA Probes; D004269:DNA, Bacterial / Q000302:isolation & purification; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005784:Gene Amplification; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid
chemical_list: D003062:Codon; D015342:DNA Probes; D004269:DNA, Bacterial; D004258:DNA Polymerase III