caa a22 4500 465822312 CHVBK 20180323112138.0 cr unu---uuuuu 170327e19900101xx s 000 0 eng 10.1007/BF00903778 doi (NATIONALLICENCE)springer-10.1007/BF00903778 Optimum conditions for efficient transformation of Streptomyces venezuelae protoplasts [Elektronische Daten] [Jozef Anné, Lieve Van Mellaert, Hendrik Eyssen] Summary: Optimum conditions for protoplast regeneration and transformation ofStreptomyces venezuelae ETH 14630 have been established. Protoplasts from mycelium grown to the stationary phase and treated with lysozyme in P medium under mild conditions gave the best regeneration frequency. Transformation of protoplasts with naked DNA was very efficient using either polyethylene glycol of mol. wt. 4000 or 6000, at concentrations of 28.5% or 36% (w/v) respectively. About 105 transformants/μg DNA could be isolated using protoplasts derived from cells cultivated to the early exponential growth phase in LB medium containing 0.2%-0.6% glycine and subsequently treated at 30°-32°C with 20 mg lysozyme/ml in P medium for 30 min. Selection of the transformants occurred on MRYE plates containing less than 105 regenerating protoplasts per plate. Higher protoplast densities considerably decreased the regeneration frequency of the transformants. Springer-Verlag, 1990 Anné Jozef Department of Microbiology, Rega Institute, University of Leuven, B-3000, Leuven, Belgium aut Van Mellaert Lieve Department of Microbiology, Rega Institute, University of Leuven, B-3000, Leuven, Belgium aut Eyssen Hendrik Department of Microbiology, Rega Institute, University of Leuven, B-3000, Leuven, Belgium aut Applied Microbiology and Biotechnology Springer-Verlag 32/4(1990-01-01), 431-435 0175-7598 32:4<431 1990 32 253 https://doi.org/10.1007/BF00903778 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00903778 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Anné Jozef Department of Microbiology, Rega Institute, University of Leuven, B-3000, Leuven, Belgium aut NATIONALLICENCE 700 1- Van Mellaert Lieve Department of Microbiology, Rega Institute, University of Leuven, B-3000, Leuven, Belgium aut NATIONALLICENCE 700 1- Eyssen Hendrik Department of Microbiology, Rega Institute, University of Leuven, B-3000, Leuven, Belgium aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer-Verlag 32/4(1990-01-01), 431-435 0175-7598 32:4<431 1990 32 253 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer