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. 1982 Oct;152(1):351–356. doi: 10.1128/jb.152.1.351-356.1982

Cloning and identification of the product of the dnaE gene of Escherichia coli.

M M Welch, C S McHenry
PMCID: PMC221416  PMID: 6288664

Abstract

We successively subcloned the dnaE gene of Escherichia coli into pBR322, resulting in a plasmid that contains 4.6 kilobases of E. coli DNA. This plasmid can complement a dnaE temperature-sensitive mutation. A restriction map of the dnaE gene and the surrounding 10.7-kilobase region of the E. coli chromosome was determined. A unique HindIII restriction endonuclease site within the cloned segment of DNA was identified as a site required for expression of the dnaE gene. By using the maxicell plasmid-directed protein synthesizing system, we demonstrated that dnaE codes for the alpha subunit of DNA polymerase III.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bendiak D. S., Friesen J. D. Organization of genes in the four minute region of the Escherichia coli chromosome: evidence that rpsB and tsf are co-transcribed. Mol Gen Genet. 1981;181(3):356–362. doi: 10.1007/BF00425611. [DOI] [PubMed] [Google Scholar]
  2. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  3. Brent R., Ptashne M. The lexA gene product represses its own promoter. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1932–1936. doi: 10.1073/pnas.77.4.1932. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burgers P. M., Kornberg A., Sakakibara Y. The dnaN gene codes for the beta subunit of DNA polymerase III holoenzyme of escherichia coli. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5391–5395. doi: 10.1073/pnas.78.9.5391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clarke L., Carbon J. A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell. 1976 Sep;9(1):91–99. doi: 10.1016/0092-8674(76)90055-6. [DOI] [PubMed] [Google Scholar]
  6. Clewell D. B., Helinski D. R. Properties of a supercoiled deoxyribonucleic acid-protein relaxation complex and strand specificity of the relaxation event. Biochemistry. 1970 Oct 27;9(22):4428–4440. doi: 10.1021/bi00824a026. [DOI] [PubMed] [Google Scholar]
  7. Clewell D. B. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol. 1972 May;110(2):667–676. doi: 10.1128/jb.110.2.667-676.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gefter M. L., Hirota Y., Kornberg T., Wechsler J. A., Barnoux C. Analysis of DNA polymerases II and 3 in mutants of Escherichia coli thermosensitive for DNA synthesis. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3150–3153. doi: 10.1073/pnas.68.12.3150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Henson J. M., Chu H., Irwin C. A., Walker J. R. Isolation and characterization of dnaX and dnaY temperature-sensitive mutants of Escherichia coli. Genetics. 1979 Aug;92(4):1041–1059. doi: 10.1093/genetics/92.4.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hübscher U., Kornberg A. The delta subunit of Escherichia coli DNA polymerase III holoenzyme is the dnaX gene product. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6284–6288. doi: 10.1073/pnas.76.12.6284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Lathe R., Lecocq J. P. The firA gene, a locus involved in the expression of rifampicin resistance in Escherichia coli. I. Characterisation of lambdafirA transducing phages constructed in vitro. Mol Gen Genet. 1977 Jul 7;154(1):43–51. doi: 10.1007/BF00265575. [DOI] [PubMed] [Google Scholar]
  13. McHenry C. S., Crow W. DNA polymerase III of Escherichia coli. Purification and identification of subunits. J Biol Chem. 1979 Mar 10;254(5):1748–1753. [PubMed] [Google Scholar]
  14. McHenry C. S. Purification and characterization of DNA polymerase III'. Identification of tau as a subunit of the DNA polymerase III holoenzyme. J Biol Chem. 1982 Mar 10;257(5):2657–2663. [PubMed] [Google Scholar]
  15. Norgard M. V., Emigholz K., Monahan J. J. Increased amplification of pBR322 plasmid deoxyribonucleic acid in Escherichia coli K-12 strains RR1 and chi1776 grown in the presence of high concentrations of nucleoside. J Bacteriol. 1979 Apr;138(1):270–272. doi: 10.1128/jb.138.1.270-272.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sakakibara Y., Tsukano H., Sako T. Organization and transcription of the dnaA and dnaN genes of Escherichia coli. Gene. 1981 Jan-Feb;13(1):47–55. doi: 10.1016/0378-1119(81)90042-1. [DOI] [PubMed] [Google Scholar]
  17. Sancar A., Hack A. M., Rupp W. D. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. doi: 10.1128/jb.137.1.692-693.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shizuya H., Livingston D. M., Richardson C. C. Isolation of a specialized transducing bacteriophage lambda carrying the polC locus of Escherichia coli. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2614–2617. doi: 10.1073/pnas.71.7.2614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Spanos A., Sedgwick S. G., Yarranton G. T., Hübscher U., Banks G. R. Detection of the catalytic activities of DNA polymerases and their associated exonucleases following SDS-polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Apr 24;9(8):1825–1839. doi: 10.1093/nar/9.8.1825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wickner S. H., Wickner R. B., Raetz C. R. Overproduction of DNA gene products by Escherichia coli strains carrying hybrid ColE1 plasmids. Biochem Biophys Res Commun. 1976 May 17;70(2):389–396. doi: 10.1016/0006-291x(76)91058-5. [DOI] [PubMed] [Google Scholar]
  21. Worcel A., Burgi E. Properties of a membrane-attached form of the folded chromosome of Escherichia coli. J Mol Biol. 1974 Jan 5;82(1):91–105. doi: 10.1016/0022-2836(74)90576-2. [DOI] [PubMed] [Google Scholar]
  22. Yamamoto M., Strycharz W. A., Nomura M. Identification of genes for elongation factor Ts and ribosomal protein S2 in E. coli. Cell. 1976 May;8(1):129–138. doi: 10.1016/0092-8674(76)90194-x. [DOI] [PubMed] [Google Scholar]

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