Summary
Polyethylene glycol (PEG) efficiently mediated the transformation ofStreptomyces avermitilis protoplasts by plasmid DNA to yield 107 transformants per μg of plasmid DNA. Under conditios in which the maximum transformation frequency was observed, the cotransformation frequency exceeded 10%. The number of transformants increased linearly with the amount of DNA and number ofS. avermitilis protoplasts. Relaxed and supercoiled, but not linear DNA transformed protoplasts efficiently. Dimethyl sulfoxide (DMSO)-mediated transformation of protoplasts was 1000-fold less efficient. PEG and, less efficiently, DMSO also mediated the transformation of whole cells ofS. avermitilis by DNA.
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Bibb, B.J., J.M. Ward and D.A. Hopwood. 1978. Transformation intoStreptomyces at high frequency. Nature 274:398–400.
Burg, R.W., B.M. Miller, E.E. Baker, J. Birnbaum, S. Currie, R. Hartman, Y.L. Kong, R.L. Monaghan, G. Olson, I. Putter, J.B. Tunac, H. Wallic, E.O. Stapley, R. Oiwa and S. Omura. 1979. Avermectins, new family of potent anthelmintic agents: producing organisms and fermentation. Antimidrob. Agents Chemother. 15: 361–367.
Fisher, M.H. and H. Mrozik. 1984. The avermectin family of macrolide-like antibiotics. In: Macrolide Antibiotics (Omura, S., ed.), pp. 553–604, Academic Press, Orlando.
Foor, F., G.P. Roberts, N. Morin, L. Snyder, M. Hwang, P.H. Gibbons, M.J. Paradiso, R.L. Stodish, C.L. Ruby, B. Wolanski and S.L. Streicher. 1985. Isolation and characterization of theStreptomyces cattleya temperate phage TG1. Gene 39: 11–16.
Greene, J. and R.B. Goldberg. 1985. Isolation and preliminary characterization of lytic and lysogenic phages with wide host range within the Streptomycetes. J. Gen. Microbiol. 131:2459–2465.
Hopwood, D.A., M.J. Bibb, K.F. Chater, T. Kieser, C.J. Burton, H.M. Kieser, D.J. Lydiate, C.P. Smith, J.M. Ward and H. Schrempf. 1985. Genetic Manipulation ofStreptomyces. A Laboratory Manual. John Innes Foundation, Norwich.
Jones, M.D., J.A. Mabe, W.M. Nakatsukasa and J.T. Fayerman. 1984. pFJ265, a new cloning vehicle forStreptomyces. Plasmid 11:92–95.
Kawai, S. and M. Nishizawa. 1984. New procedure for DNA transfection with polycation and dimethyl sulfoxide. Mol. Cell. Biol. 4:1172–1174.
Kieser, T., D.A. Hopwood, H.M. Wright and C.J. Thompson. 1982. pIJ101, a multi-copy broad host-rangeStreptomyces plasmid: functional analysis and development of DNA cloning vectors. Mol. Gen. Genet. 185: 223–228.
Konvalinkova, V., P. Roelants and M. Mergeay. 1977. Transfection inStreptomyces virginiae. Biochem. Soc. Trans. 5: 941–943.
Lampel, J.S. and W.R. Strohl. 1986. Transformation and transfection of anthracycline-producingStreptomyces. Appl. Environ. Microbiol. 51: 126–131.
MacNeil, D.J. 1986. A flexible boiling procedure for isolating plasmid DNA from gram-positive microorganisms. J. Microbiol. Methods 5: 115–124.
MacNeil, T. and P.H. Gibbons. 1986. Characterization of theStreptomyces plasmid pVE1. Plasmid 16: 182–194.
Matsushima, P. and R.H. Baltz. 1985. Efficient plasmid transformation ofStreptomyces ambofaciens andStreptomyces fradiae protoplasts. J. Bacteriol. 163: 180–185.
Ogata, S., Y. Koyama-Miyosh and S. Hayashida. 1985. Transfection and transformation systems for pock-forming and thiostrepton-producingStreptomyces azureus. J. Fac. Agric. Kyushu Univ. 29: 179–188.
Okanishi, M., K. Suzuki and H. Umezawa. 1974. Formation and reversion of Streptomycete protoplasts: culture conditions and morphological study. J. Gen. Microbiol. 80: 389–400.
Petricek, M., I. Smrckova and P. Tichy. 1985. Transformation ofStreptomyces granaticolor with natural and recombinant plasmid vectors. Folia Microbiol. 30: 474–478.
Putter, I., J.G. MacConnell, F.A. Preiser, A.A. Haidri, S.S. Ristich and R.A. Dybas. 1981. Avermectins: novel insecticides, acaricides and nematicides from a soil microorganism. Experientia 37: 963–964.
Rodicio, M.R., C.J. Burton and K.F. Chater. 1985. New derivatives of theStreptomyces temperate phage ØC31 useful for the cloning and functional analysis ofStreptomyces DNA. Gene 34: 283–292.
Rodicio, M.R. and K.F. Chater. 1982. Small DNA-free liposomes stimulate transfection ofStreptomyces protoplasts. J. Bacteriol. 142: 8–14.
Suarez, J.E. and K.F. Chater. 1980. Polyethylene glycol-assisted transfection ofStreptomyces protoplasts. J. Bacteriol. 142: 8–14.
Thompson, C.J., J.M. Ward and D.A. Hopwood. 1982. Cloning of antibiotic resistance and nutritional genes in Streptomycetes. J. Bacteriol. 151: 668–677.
Zenghui, Z., T. Huarong, X. Yi, Z. Yingfan, and X. Yugu. 1980. A study on transformation ofStreptomyces griseus plasmid DNA. Acta Genet. Sinica 7: 291–298.
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MacNeil, D.J., Klapko, L.M. Transformation ofStreptomyces avermitilis by plasmid DNA. Journal of Industrial Microbiology 2, 209–218 (1987). https://doi.org/10.1007/BF01569542
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DOI: https://doi.org/10.1007/BF01569542