Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1988 May 25;16(10):4341–4352. doi: 10.1093/nar/16.10.4341

Site-specific degradation of Streptomyces lividans DNA during electrophoresis in buffers contaminated with ferrous iron.

X Zhou 1, Z Deng 1, J L Firmin 1, D A Hopwood 1, T Kieser 1
PMCID: PMC336634  PMID: 2837731

Abstract

Streptomyces lividans DNA contains a modification which makes it susceptible to double-strand cleavage during electrophoresis in buffers contaminated with ferrous iron (which may be present in some batches of EDTA). The cleavage of the DNA is site-specific and the average fragment size resulting from limit digestion of total S. lividans DNA is about 6kb. DNA from Streptomyces coelicolor A3(2) and several other Streptomyces strains, and from E. coli, is not cleaved under the same conditions. A S. lividans mutant has been isolated which lacks the DNA modification. We suspect that many reports of "poor" preparations of S. lividans plasmids may be due to the above effect.

Full text

PDF
4341

Images in this article

4343Image
on p.4343
4345Image
on p.4345
4346Image
on p.4346
4348Image
on p.4348
4349Image
on p.4349

Selected References

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

  1. Altenbuchner J., Cullum J. DNA amplification and an unstable arginine gene in Streptomyces lividans 66. Mol Gen Genet. 1984;195(1-2):134–138. doi: 10.1007/BF00332735. [DOI] [PubMed] [Google Scholar]
  2. Chater K. F., Wilde L. C. Streptomyces albus G mutants defective in the SalGI restriction-modification system. J Gen Microbiol. 1980 Feb;116(2):323–334. doi: 10.1099/00221287-116-2-323. [DOI] [PubMed] [Google Scholar]
  3. Dervan P. B. Design of sequence-specific DNA-binding molecules. Science. 1986 Apr 25;232(4749):464–471. doi: 10.1126/science.2421408. [DOI] [PubMed] [Google Scholar]
  4. Hertzberg R. P., Dervan P. B. Cleavage of DNA with methidiumpropyl-EDTA-iron(II): reaction conditions and product analyses. Biochemistry. 1984 Aug 14;23(17):3934–3945. doi: 10.1021/bi00312a022. [DOI] [PubMed] [Google Scholar]
  5. Hopwood D. A., Kieser T., Wright H. M., Bibb M. J. Plasmids, recombination and chromosome mapping in Streptomyces lividans 66. J Gen Microbiol. 1983 Jul;129(7):2257–2269. doi: 10.1099/00221287-129-7-2257. [DOI] [PubMed] [Google Scholar]
  6. Kan N. C., Lautenberger J. A., Edgell M. H., Hutchison C. A., 3rd The nucleotide sequence recognized by the Escherichia coli K12 restriction and modification enzymes. J Mol Biol. 1979 May 15;130(2):191–209. doi: 10.1016/0022-2836(79)90426-1. [DOI] [PubMed] [Google Scholar]
  7. Kessler C., Höltke H. J. Specificity of restriction endonucleases and methylases--a review. Gene. 1986;47(1):1–153. doi: 10.1016/0378-1119(86)90245-3. [DOI] [PubMed] [Google Scholar]
  8. Kieser T. Factors affecting the isolation of CCC DNA from Streptomyces lividans and Escherichia coli. Plasmid. 1984 Jul;12(1):19–36. doi: 10.1016/0147-619x(84)90063-5. [DOI] [PubMed] [Google Scholar]
  9. Kieser T., Hopwood D. A., Wright H. M., Thompson C. J. pIJ101, a multi-copy broad host-range Streptomyces plasmid: functional analysis and development of DNA cloning vectors. Mol Gen Genet. 1982;185(2):223–228. doi: 10.1007/BF00330791. [DOI] [PubMed] [Google Scholar]
  10. Kieser T., Melton R. E. Plasmid pIJ699, a multi-copy positive-selection vector for Streptomyces. Gene. 1988 May 15;65(1):83–91. doi: 10.1016/0378-1119(88)90419-2. [DOI] [PubMed] [Google Scholar]
  11. Luckow B., Renkawitz R., Schütz G. A new method for constructing linker scanning mutants. Nucleic Acids Res. 1987 Jan 26;15(2):417–429. doi: 10.1093/nar/15.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  13. Moser H. E., Dervan P. B. Sequence-specific cleavage of double helical DNA by triple helix formation. Science. 1987 Oct 30;238(4827):645–650. doi: 10.1126/science.3118463. [DOI] [PubMed] [Google Scholar]
  14. Murray N. E., Brammar W. J., Murray K. Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet. 1977 Jan 7;150(1):53–61. doi: 10.1007/BF02425325. [DOI] [PubMed] [Google Scholar]
  15. Tsai J. F., Chen C. W. Isolation and characterization of Streptomyces lividans mutants deficient in intraplasmid recombination. Mol Gen Genet. 1987 Jun;208(1-2):211–218. doi: 10.1007/BF00330444. [DOI] [PubMed] [Google Scholar]
  16. de Vries E., Bloemers S. M., van der Vliet P. C. Incorporation of 5-bromodeoxycytidine in the adenovirus 2 replication origin interferes with nuclear factor 1 binding. Nucleic Acids Res. 1987 Sep 25;15(18):7223–7234. doi: 10.1093/nar/15.18.7223. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES