Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1991 Mar;57(3):665–671. doi: 10.1128/aem.57.3.665-671.1991

Construction of a hybrid plasmid capable of replication in Amycolatopsis mediterranei.

R Lal 1, S Lal 1, E Grund 1, R Eichenlaub 1
PMCID: PMC182777  PMID: 2039229

Abstract

A new plasmid, pA387, has been isolated from "Amycolatopsis sp." (DSM 43387). This plasmid could be isolated from liquid culture as well as mycelium from agar plates by a modified procedure. Plasmid pA387 is about 29.6 kb and can be cured at low frequency by protoplasting and ethidium bromide and heat treatment. Hybridization experiments showed that this plasmid is present in free form and does not integrate into the chromosome. A hybrid plasmid was constructed by cloning a 5.1-kb fragment of pA387 into the Escherichia coli vector pDM10. This hybrid plasmid, termed pRL1, could be transformed into Amycolatopsis mediterranei and A. orientalis by electroporation. A transformation frequency of 2.2 x 10(3) transformants per micrograms of DNA at 12.5 kV/cm and a pulse duration of 10.8 ms was obtained in A. mediterranei, whereas 1.1 x 10(5) transformants per microgram of DNA were obtained at a field strength of 7.5 kV/cm and a pulse duration of 7.6 ms in A. orientalis. Plasmid pRL1 is the first hybrid plasmid which could be used successfully for the transformation of A. mediterranei. The plasmid has a rather high copy number, is genetically stable, and can be easily reisolated from A. mediterranei. Plasmid pRL1 will be useful for further construction of a shuttle vector for E. coli and A. mediterranei and becomes the basis for the development of gene cloning techniques in Amycolatopsis spp.

Full text

PDF
665

Selected References

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

  1. Allen S. P., Blaschek H. P. Electroporation-induced transformation of intact cells of Clostridium perfringens. Appl Environ Microbiol. 1988 Sep;54(9):2322–2324. doi: 10.1128/aem.54.9.2322-2324.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barna J. C., Williams D. H. The structure and mode of action of glycopeptide antibiotics of the vancomycin group. Annu Rev Microbiol. 1984;38:339–357. doi: 10.1146/annurev.mi.38.100184.002011. [DOI] [PubMed] [Google Scholar]
  3. Bibb M. J., Ward J. M., Hopwood D. A. Transformation of plasmid DNA into Streptomyces at high frequency. Nature. 1978 Jul 27;274(5669):398–400. doi: 10.1038/274398a0. [DOI] [PubMed] [Google Scholar]
  4. Crameri R., Davies J. E., Hütter R. Plasmid curing and generation of mutations induced with ethidium bromide in streptomycetes. J Gen Microbiol. 1986 Mar;132(3):819–824. doi: 10.1099/00221287-132-3-819. [DOI] [PubMed] [Google Scholar]
  5. Dower W. J., Miller J. F., Ragsdale C. W. High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res. 1988 Jul 11;16(13):6127–6145. doi: 10.1093/nar/16.13.6127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Eckhardt T. A rapid method for the identification of plasmid desoxyribonucleic acid in bacteria. Plasmid. 1978 Sep;1(4):584–588. doi: 10.1016/0147-619x(78)90016-1. [DOI] [PubMed] [Google Scholar]
  7. Grund E., Knorr C., Eichenlaub R. Catabolism of benzoate and monohydroxylated benzoates by Amycolatopsis and Streptomyces spp. Appl Environ Microbiol. 1990 May;56(5):1459–1464. doi: 10.1128/aem.56.5.1459-1464.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hattermann D. R., Stacey G. Efficient DNA transformation of Bradyrhizobium japonicum by electroporation. Appl Environ Microbiol. 1990 Apr;56(4):833–836. doi: 10.1128/aem.56.4.833-836.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hopwood D. A., Bibb M. J., Chater K. F., Kieser T. Plasmid and phage vectors for gene cloning and analysis in Streptomyces. Methods Enzymol. 1987;153:116–166. doi: 10.1016/0076-6879(87)53052-x. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Kim A. Y., Blaschek H. P. Construction of an Escherichia coli-Clostridium perfringens shuttle vector and plasmid transformation of Clostridium perfringens. Appl Environ Microbiol. 1989 Feb;55(2):360–365. doi: 10.1128/aem.55.2.360-365.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. MacNeil D. J. Characterization of a unique methyl-specific restriction system in Streptomyces avermitilis. J Bacteriol. 1988 Dec;170(12):5607–5612. doi: 10.1128/jb.170.12.5607-5612.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Madon J., Moretti P., Hütter R. Site-specific integration and excision of pMEA100 in Nocardia mediterranei. Mol Gen Genet. 1987 Sep;209(2):257–264. doi: 10.1007/BF00329651. [DOI] [PubMed] [Google Scholar]
  14. Matsushima P., Baltz R. H. Efficient plasmid transformation of Streptomyces ambofaciens and Streptomyces fradiae protoplasts. J Bacteriol. 1985 Jul;163(1):180–185. doi: 10.1128/jb.163.1.180-185.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Matsushima P., McHenney M. A., Baltz R. H. Efficient transformation of Amycolatopsis orientalis (Nocardia orientalis) protoplasts by Streptomyces plasmids. J Bacteriol. 1987 May;169(5):2298–2300. doi: 10.1128/jb.169.5.2298-2300.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Meletzus D., Eichenlaub R. Transformation of the phytopathogenic bacterium Clavibacter michiganense subsp. michiganense by electroporation and development of a cloning vector. J Bacteriol. 1991 Jan;173(1):184–190. doi: 10.1128/jb.173.1.184-190.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Miller J. F., Dower W. J., Tompkins L. S. High-voltage electroporation of bacteria: genetic transformation of Campylobacter jejuni with plasmid DNA. Proc Natl Acad Sci U S A. 1988 Feb;85(3):856–860. doi: 10.1073/pnas.85.3.856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moretti P., Hintermann G., Hütter R. Isolation and characterization of an extrachromosomal element from Nocardia mediterranei. Plasmid. 1985 Sep;14(2):126–133. doi: 10.1016/0147-619x(85)90072-1. [DOI] [PubMed] [Google Scholar]
  19. Neesen K., Volckaert G. Construction and shuttling of novel bifunctional vectors for Streptomyces spp. and Escherichia coli. J Bacteriol. 1989 Mar;171(3):1569–1573. doi: 10.1128/jb.171.3.1569-1573.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Oh Y. K., Fare L. R., Taylor D. P., Widger J., Nisbet L. J. A cryptic plasmid from Nocardia orientalis NRRL 2452, a vancomycin producer. J Antibiot (Tokyo) 1986 May;39(5):694–698. doi: 10.7164/antibiotics.39.694. [DOI] [PubMed] [Google Scholar]
  21. Powell Ian B., Achen Marc G., Hillier Alan J., Davidson Barrie E. A Simple and Rapid Method for Genetic Transformation of Lactic Streptococci by Electroporation. Appl Environ Microbiol. 1988 Mar;54(3):655–660. doi: 10.1128/aem.54.3.655-660.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schupp T., Hutter R., Hopwood D. A. Genetic recombination in Nocardia mediterranei. J Bacteriol. 1975 Jan;121(1):128–136. doi: 10.1128/jb.121.1.128-136.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Thompson C. J., Ward J. M., Hopwood D. A. Cloning of antibiotic resistance and nutritional genes in streptomycetes. J Bacteriol. 1982 Aug;151(2):668–677. doi: 10.1128/jb.151.2.668-677.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wohlleben W., Pühler A. The Streptomyces ghanaensis low copy plasmid pSG2 and its use for vector construction. Arch Microbiol. 1987 Oct;148(4):298–304. doi: 10.1007/BF00456708. [DOI] [PubMed] [Google Scholar]
  25. Yamamoto H., Maurer K. H., Hutchinson C. R. Transformation of Streptomyces erythraeus. J Antibiot (Tokyo) 1986 Sep;39(9):1304–1313. doi: 10.7164/antibiotics.39.1304. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES