Isolation, sequencing and overexpression of the gene encoding the theta subunit of DNA polymerase III holoenzyme.
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.
Carter
J R
JR
University of Colorado Health Sciences Center, Department of Biochemistry, Biophysics and Genetics, Denver 80262.
Franden
M A
MA
Aebersold
R
R
Kim
D R
DR
McHenry
C S
CS
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
D003062:Codon; D015342:DNA Probes; D004269:DNA, Bacterial; D004258:DNA Polymerase III