PMID:8509334
| Citation |
Carter, JR, Franden, MA, Aebersold, R and McHenry, CS (1993) Identification, isolation, and characterization of the structural gene encoding the delta' subunit of Escherichia coli DNA polymerase III holoenzyme. J. Bacteriol. 175:3812-22 |
|---|---|
| Abstract |
The gene encoding the delta' subunit of DNA polymerase III holoenzyme, designated holB, was cloned by a strategy in which peptide sequence was used to derive a DNA hybridization probe. The gene maps to 24.95 centisomes of the chromosome. Sequencing of holB revealed a 1,002-bp open reading frame predicted to produce a 36,936-Da protein. The gene has a ribosome-binding site and promoter that are highly similar to the consensus sequences and is flanked by two potential open reading frames. Protein sequence analysis of delta' revealed a high degree of similarity to the dnaX gene products of Escherichia coli and Bacillus subtilis, including one stretch of 10 identical amino acid residues. A lesser degree of similarity to the gene 44 protein of bacteriophage T4 and the 40-kDa protein of the A1 complex (replication factor C) of HeLa cells was seen. The gene, when placed into a tac promoter-based expression plasmid, directed expression of two proteins of similar size. By immunodetection with anti-holoenzyme immunoglobulin G, both proteins are judged to be products of holB. |
| Links | |
| Keywords |
Amino Acid Sequence; Bacterial Proteins/genetics; Base Sequence; Chromosome Mapping; Cloning, Molecular; Codon; DNA Polymerase III/genetics; DNA, Bacterial/genetics; Escherichia coli/enzymology; Escherichia coli/genetics; Genes, Bacterial; Molecular Sequence Data; Oligodeoxyribonucleotides/chemistry; Peptide Fragments/chemistry; Polymerase Chain Reaction; Recombinant Proteins/metabolism; Restriction Mapping |
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