Identification, isolation, and characterization of the structural gene encoding the delta' subunit of Escherichia coli DNA polymerase III holoenzyme. Journal Article uri icon

Overview

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.

publication date

  • June 1, 1993

Date in CU Experts

  • October 1, 2013 12:09 PM

Full Author List

  • Carter JR; Franden MA; Aebersold R; McHenry CS

author count

  • 4

citation count

  • 9

Other Profiles

International Standard Serial Number (ISSN)

  • 0021-9193

Electronic International Standard Serial Number (EISSN)

  • 1098-5530

Additional Document Info

start page

  • 3812

end page

  • 3822

volume

  • 175

issue

  • 12