Home LiteratureArticle Details
PMID: 372736 Published · ppublish English Journal Article

Plasmid replication functions. II. Cloning analysis of the repA replication region of antibiotic resistance plasmid R6-5.

Molecular & general genetics : MGG ·Vol. 168 ·No. 1 ·1979-01-05 ·Pages 1-25

Andrés I, Slocombe PM, Cabello F, Timmis JK, Lurz R, Burkardt HJ, Timmis KN

Abstract

R6-5 is a low copy number, conjugative, FII incompatibility group plasmid that has a molecular length of 102 kb and that specifies resistance against several antibiotics (chloramphenicol, fusidic acid, kanamycin, streptomycin and sulphonamide) and mercury salts. By means of in vitro cloning procedures, mini plasmids have been generated that contain a DNA segment from the essential region of R6-5 that is only 2.6 kb in length. This DNA segment, which consists of two PstI fragments that are adjacent in the parent plasmid, carries all genes and sequences required for the regulated replication and incompatibility properties of R6-5, including its origin of replication, OriV, an essential function that has been designated RepA, and the copy control function, Cop. Three different polypeptides, having monomer molecular weights of 23,000, 10,000 and 9,500 daltons, are synthesized in detectable quantities by minicells carrying pBR322 hybrid plasmids that contain DNA segments from the R6-5 essential region. A spontaneous deletion derivative of a pBR322 hybrid plasmid that carries the R6-5 origin of replication was isolated. Heteroduplex analysis of this derivative plasmid indicates that the deleted DNA segment carries the R6-5 replication origin and that its termini consist of short inverted repeat sequences.

MeSH Terms
DNA Polymerase I/metabolism DNA Replication DNA Restriction Enzymes/metabolism DNA, Bacterial/metabolism DNA, Recombinant/metabolism Electrophoresis, Agar Gel Escherichia coli/genetics Genes Phenotype R Factors Transformation, Bacterial
Chemicals
DNA, Bacterial DNA, Recombinant DNA Polymerase I DNA Restriction Enzymes
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Andrés I
Slocombe P M
Cabello F
Timmis J K
Lurz R
Burkardt H J
Timmis K N
References (51)
51 references, click to expand
  1. Construction of biologically functional bacterial plasmids in vitro.
    Proc Natl Acad Sci U S A. 1973 Nov;70(11):3240-4 PMID: 4594039
  2. Phage lambda receptor chromosomes for DNA fragments made with restriction endonuclease III of Haemophilus influenzae and restriction endonuclease I of Escherichia coli.
    J Mol Biol. 1975 Nov 5;98(3):551-64 PMID: 1104875
  3. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form.
    Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159-66 PMID: 4894690
  4. Genome construction between bacterial species in vitro: replication and expression of Staphylococcus plasmid genes in Escherichia coli.
    Proc Natl Acad Sci U S A. 1974 Apr;71(4):1030-4 PMID: 4598290
  5. Replication of colicin E1 plasmid DNA added to cell extracts.
    Proc Natl Acad Sci U S A. 1975 Mar;72(3):1050-4 PMID: 1093162
  6. Plasmid-encoded functions involved in the replication and inheritance of antibiotic-resistance plasmid R6-5.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:105-10 PMID: 383374
  7. Construction of a novel plasmid-phage hybrid: use of the hybrid to demonstrate ColE1 DNA replication in vivo in the absence of a ColE1-specified protein.
    Proc Natl Acad Sci U S A. 1978 May;75(5):2200-4 PMID: 276860
  8. Plasmid incompatibility and control of replication: copy mutants of the R-factor R1 in Escherichia coli K-12.
    J Bacteriol. 1975 Nov;124(2):641-9 PMID: 1102525
  9. A simple method for the preparation of large quantities of pure plasmid DNA.
    Biochim Biophys Acta. 1975 Apr 2;383(4):457-63 PMID: 1092355
  10. Molecular cloning of DNA from F sex factor of Escherichia coli K-12.
    Proc Natl Acad Sci U S A. 1976 Jan;73(1):64-8 PMID: 1108020
  11. Conjugational complementation analysis of transfer-deficient mutants of Flac in Escherichia coli.
    J Bacteriol. 1972 Jun;110(3):831-42 PMID: 4555412
  12. A routine method for protein-free spreading of double- and single-stranded nucleic acid molecules.
    Proc Natl Acad Sci U S A. 1975 Jan;72(1):83-7 PMID: 164030
  13. Cloning, isolation, and characterization of replication regions of complex plasmid genomes.
    Proc Natl Acad Sci U S A. 1975 Jun;72(6):2242-6 PMID: 1094460
  14. The transposon Tn1 as a probe for studying ColE1 structure and function.
    Mol Gen Genet. 1977 Mar 7;151(2):151-60 PMID: 327263
  15. 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
  16. Isolation of large molecular weight DNA from agarose gels for further digestion by restriction enzymes.
    FEBS Lett. 1975 Apr 15;53(1):84-6 PMID: 1140401
  17. Nonchromosomal antibiotic resistance in bacteria. V. Isolation and characterization of R factor mutants exhibiting temperature-sensitive repression of fertility.
    J Bacteriol. 1972 Jun;110(3):1082-8 PMID: 4555403
  18. Unidirectional replication in Escherichia coli of three small plasmids derived from R factor R12.
    Plasmid. 1977 Nov;1(1):8-18 PMID: 375272
  19. Restriction endonucleases.
    CRC Crit Rev Biochem. 1976 Nov;4(2):123-64 PMID: 795607
  20. Cloning and characterization of EcoRI and HindIII restriction endonuclease-generated fragments of antibiotic resistance plasmids R6-5 and R6.
    Mol Gen Genet. 1978 Jun 14;162(2):121-37 PMID: 672900
  21. Molecular cloning of an Escherichia coli plasmid determinant than encodes for the production of heat-stable enterotoxin.
    J Bacteriol. 1976 Oct;128(1):463-72 PMID: 789348
  22. Class of small multicopy plasmids originating from the mutant antibiotic resistance factor R1 drd-19B2.
    J Bacteriol. 1975 Aug;123(2):658-65 PMID: 1097421
  23. Plasmid ColE1 DNA replication in Escherichia coli strains temperature-sensitive for DNA replication.
    Mol Gen Genet. 1975;136(4):273-89 PMID: 16094996
  24. Method for the isolation of the replication region of a bacterial replicon: construction of a mini-F'kn plasmid.
    J Bacteriol. 1976 Aug;127(2):982-7 PMID: 783124
  25. F plasmid incompatibility and copy number genes: their map locations and interactions.
    Plasmid. 1978 Sep;1(4):492-507 PMID: 372978
  26. DNA polymerase as a requirement for the maintenance of the bacterial plasmid colicinogenic factor E1.
    Biochem Biophys Res Commun. 1970 Dec 24;41(6):1538-44 PMID: 4922634
  27. Isolation and characterization of the minimal fragment required for autonomous replication ("basic replicon") of a copy mutant (pKN102) of the antibiotic resistance factor R1.
    Mol Gen Genet. 1978 Jun 1;162(1):51-7 PMID: 353525
  28. Fertility repression of F-like conjugative plasmids: physical mapping of the R6--5 finO and finP cistrons and identification of the finO protein.
    Proc Natl Acad Sci U S A. 1978 Dec;75(12):5836-40 PMID: 366604
  29. Temperature-sensitive mutants for the replication of plasmids in Escherichia coli: requirement for deoxyribonucleic acid polymerase I in the replication of the plasmid ColE 1 .
    J Bacteriol. 1973 Jun;114(3):1116-24 PMID: 4576398
  30. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels.
    J Mol Biol. 1977 Feb 15;110(1):119-46 PMID: 845942
  31. Replication control in a composite plasmid constructed by in vitro linkage of two distinct replicons.
    Nature. 1976 Jan 29;259(5541):285-90 PMID: 765836
  32. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.
    Gene. 1977;2(2):95-113 PMID: 344137
  33. Molecular cloning of DNA fragments produced by restriction endonucleases Sa1I and BamI.
    Proc Natl Acad Sci U S A. 1976 May;73(5):1537-41 PMID: 818638
  34. Plasmid mediated alterations in composition and structure of envelopes of Escherichia coli B/r.
    Biochim Biophys Acta. 1977 Oct 17;470(2):258-72 PMID: 334260
  35. Instability of plasmid DNA sequences: macro and micro evolution of the antibiotic resistance plasmid R6-5.
    Mol Gen Genet. 1978 Nov 16;167(1):11-19 PMID: 368583
  36. Plasmid replication functions. III. Origin and direction of replication of a "mini" plasmid derived from R6-5.
    Mol Gen Genet. 1979 Jan 5;168(1):27-36 PMID: 372738
  37. Covalently closed circular DNA molecules of low superhelix density as intermediate forms in plasmid replication.
    Nature. 1976 Jun 10;261(5560):512-6 PMID: 778630
  38. The control region of the F sex factor DNA transfer cistrons: restriction mapping and DNA cloning.
    Mol Gen Genet. 1978 Oct 24;165(3):295-304 PMID: 368564
  39. Replication region fragments cloned from Flac+ are identical to EcoRI fragment f5 of F.
    J Bacteriol. 1976 Sep;127(3):1571-5 PMID: 956131
  40. Pedigrees of some mutant strains of Escherichia coli K-12.
    Bacteriol Rev. 1972 Dec;36(4):525-57 PMID: 4568763
  41. Immunological studies of aminoglycoside 3'-phosphotransferases.
    J Antibiot (Tokyo). 1976 Oct;29(10):1127-8 PMID: 825667
  42. Utilization of two distinct modes of replication by a hybrid plasmid constructed in vitro from separate replicons.
    Proc Natl Acad Sci U S A. 1974 Nov;71(11):4556-60 PMID: 4612523
  43. Mating aggregates in Escherichia coli conjugation.
    J Bacteriol. 1975 Aug;123(2):505-15 PMID: 1097414
  44. Bidirection replication from a unique origin in a mini-F plasmid.
    Proc Natl Acad Sci U S A. 1977 Mar;74(3):1138-41 PMID: 322144
  45. Conjugation proteins encoded by the F sex factor.
    Nature. 1977 Dec 15;270(5638):580-5 PMID: 339109
  46. Analysis of bacteriophage T7 early RNAs and proteins on slab gels.
    J Mol Biol. 1973 Sep 15;79(2):237-48 PMID: 4760132
  47. Plasmid incompatibility: cloning analysis of an incFII determinant of R6-5.
    Nature. 1978 May 4;273(5657):27-32 PMID: 692666
  48. Identification and mapping of the replication genes of an R factor, R100-1, integrated into the chromosome of Escherichia coli K-12.
    J Bacteriol. 1974 Jun;118(3):1123-31 PMID: 4597997
  49. Translation of Drosophila melanogaster sequences in Escherichia coli.
    Proc Natl Acad Sci U S A. 1977 Nov;74(11):5041-5 PMID: 412196
  50. Replication of colicin E1 plasmid DNA in vivo requires no plasmid-encoded proteins.
    J Bacteriol. 1978 Mar;133(3):1287-94 PMID: 346567
  51. Isolation, by tetracycline selection, of small plasmids derived from R-factor R12 in Escherichia coli K-12.
    J Bacteriol. 1976 Jul;127(1):644-55 PMID: 776944
Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00267929
Published
1979-01-05
Pages
1-25
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00267929
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: [email protected]

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