Home LiteratureArticle Details
PMID: 7009552 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Construction and characterization of hybrid plasmids containing the Escherichia coli nrd region.

Journal of bacteriology ·Vol. 143 ·No. 2 ·1980-08-00 ·Pages 561-8

Platz A, Sjöberg BM

Abstract

Recombinant plasmids containing all or part of the genetic region of Escherichia coli coding for the two subunits of ribonucleoside diphosphate reductase (proteins B1 and B2) were constructed with the aid of the multicopy plasmid pBR322. Two of these plasmids (pPS1 and pPS2) appeared to carry both a regulator and the complete structural information for the enzyme and, after transformation of E. coli, directed a 10- to 20-fold overproduction of both proteins B1 and B2. The other plasmids (pPS101 and pPS201) carried structural information for only protein B2. Cells carrying pPS1 and pPS2 showed a 5- to 500-fold increased resistance against the drug hydroxyurea. This establishes that in E. coli the inhibition of deoxyribonucleic acid synthesis by hydroxyurea is fully explained by its action on ribonucleotide reductase.

MeSH Terms
DNA, Recombinant Enzyme Induction Escherichia coli/genetics Genes Hydroxyurea/pharmacology Macromolecular Substances Plasmids Ribonucleoside Diphosphate Reductase/antagonists & inhibitors,genetics Ribonucleotide Reductases/genetics
Chemicals
DNA, Recombinant Macromolecular Substances Ribonucleotide Reductases Ribonucleoside Diphosphate Reductase Hydroxyurea
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Platz A
Sjöberg B M
References (33)
33 references, click to expand
  1. Studies with hydroxyurea. I. The reversible inhibition of bacterial DNA synthesis and the effect of hydroxyurea on the bactericidal action of streptomycin.
    Biochim Biophys Acta. 1966 Mar 21;114(3):501-15 PMID: 5330658
  2. Genetic control of L-alpha-glycerophosphate system in Escherichia coli.
    J Mol Biol. 1968 Feb 14;31(3):371-87 PMID: 4866330
  3. Inhibition of ribonucleoside diphosphate reductase by hydroxyurea.
    Cancer Res. 1968 Aug;28(8):1559-65 PMID: 4876978
  4. Ribonucleoside diphosphate reductase. Purification of the two subunits, proteins B1 and B2.
    Eur J Biochem. 1969 Jul;9(4):561-73 PMID: 4896737
  5. Turnover of the deoxyribonucleoside triphosphates in Escherichia coli 15 T during thymine starvation.
    Eur J Biochem. 1971 May 11;20(1):36-43 PMID: 4931186
  6. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol.
    J Bacteriol. 1972 May;110(2):667-76 PMID: 4336693
  7. Electron spin resonance of the iron-containing protein B2 from ribonucleotide reductase.
    J Biol Chem. 1972 Jun 10;247(11):3485-8 PMID: 4337857
  8. Defective gene product in dnaF mutant of Escherichia coli.
    Nat New Biol. 1972 Jul 19;238(81):69-71 PMID: 4558262
  9. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA.
    Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110-4 PMID: 4559594
  10. A mutant of Escherichia coli defective in ribonucleosidediphosphate reductase. 2. Characterization of the enzymatic defect.
    Eur J Biochem. 1973 Feb 1;32(3):457-62 PMID: 4571066
  11. Physicochemical characterization of ribonucleoside diphosphate reductase from Escherichia coli.
    J Biol Chem. 1973 Jul 10;248(13):4591-601 PMID: 4578086
  12. General method for the isolation of plasmid deoxyribonucleic acid.
    J Bacteriol. 1973 Nov;116(2):1064-6 PMID: 4583233
  13. Properties of a membrane-attached form of the folded chromosome of Escherichia coli.
    J Mol Biol. 1974 Jan 5;82(1):91-105 PMID: 4594427
  14. Defective bacteriophage lambda chromosome, potential vector for DNA fragments obtained after cleavage by Bacillus amyloliquefaciens endonuclease (BamI).
    FEBS Lett. 1975 Aug 1;56(1):7-11 PMID: 1157934
  15. Studies on the cleavage of bacteriophage lambda DNA with EcoRI Restriction endonuclease.
    J Mol Biol. 1975 Jan 25;91(3):315-28 PMID: 1102702
  16. Location in bacteriophage lamdba DNA of cleavage sites of the site-specific endonuclease from Bacillus amyloliquefaciens H.
    J Virol. 1976 May;18(2):659-63 PMID: 1083914
  17. Mapping of nrdA and nrdB in Escherichia coli K-12.
    J Bacteriol. 1976 Dec;128(3):810-4 PMID: 791934
  18. Gene cloning and containment properties of plasmid Col E1 and its derivatives.
    Science. 1977 Apr 8;196(4286):172-4 PMID: 322277
  19. Rat insulin genes: construction of plasmids containing the coding sequences.
    Science. 1977 Jun 17;196(4296):1313-9 PMID: 325648
  20. Ribonucleoside diphosphate reductase from Escherichia coli. An immunological assay and a novel purification from an overproducing strain lysogenic for phage lambdadnrd.
    J Biol Chem. 1977 Sep 10;252(17):6132-8 PMID: 197082
  21. Mechanism of action of nalidixic acid: purification of Escherichia coli nalA gene product and its relationship to DNA gyrase and a novel nicking-closing enzyme.
    Proc Natl Acad Sci U S A. 1977 Nov;74(11):4767-71 PMID: 200930
  22. Nalidixic acid resistance: a second genetic character involved in DNA gyrase activity.
    Proc Natl Acad Sci U S A. 1977 Nov;74(11):4772-6 PMID: 337300
  23. Construction and characterization of new cloning vehicles. I. Ampicillin-resistant derivatives of the plasmid pMB9.
    Gene. 1977;2(2):75-93 PMID: 344136
  24. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.
    Gene. 1977;2(2):95-113 PMID: 344137
  25. The tyrosine free radical in ribonucleotide reductase from Escherichia coli.
    J Biol Chem. 1978 Oct 10;253(19):6863-5 PMID: 211133
  26. Simple method for identification of plasmid-coded proteins.
    J Bacteriol. 1979 Jan;137(1):692-3 PMID: 368040
  27. Transformation of mammalian cells with genes from procaryotes and eucaryotes.
    Cell. 1979 Apr;16(4):777-85 PMID: 222468
  28. Ribonucleotide reductase from calf thymus. Purification and properties.
    Biochemistry. 1979 Jul 10;18(14):2941-8 PMID: 111707
  29. Reduction of ribonucleotides.
    Annu Rev Biochem. 1979;48:133-58 PMID: 382982
  30. Use of the lambda phage promoter PL to promote gene expression in hybrid plasmid cloning vehicles.
    Methods Enzymol. 1979;68:482-92 PMID: 232223
  31. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  32. Hybridization between Escherichia coli and Shigella.
    J Bacteriol. 1957 Oct;74(4):461-76 PMID: 13475269
  33. Mutants of Escherichia coli requiring methionine or vitamin B12.
    J Bacteriol. 1950 Jul;60(1):17-28 PMID: 15436457
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.143.2.561-568.1980
Published
1980-08-00
Pages
561-8
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC294315
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.143.2.561-568.1980 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2