A Simple and Rapid Method of Transformation of Streptomyces rimosus R6 and Other Streptomycetes by Electroporation.

Applied and Environmental Microbiology
J Pigac, H Schrempf

Abstract

Usually plasmid DNA is introduced into Streptomyces strains by polyethylene glycol-mediated transformation of protoplasts. However, many Streptomyces strains are only poorly or not at all transformable via protoplasts. Therefore, we have optimized the parameters critical for the application of electrotransformation of plasmid DNA into Streptomyces species. The most critical parameters evaluated for electrotransformation of the model strain Streptomyces rimosus R6 were the pretreatment of mycelia, buffer composition, and electric field strength. The electrocompetent mycelia were prepared from 24-h-old cultures, treated mildly with lysozyme, resuspended in sucrose-glycerol-polyethylene glycol buffer, and stored in aliquots at -70 deg C. The electric field strength of 10 kV/cm at 400 (Omega) and a capacitance of 25 (mu)F was applied. The method is simple and rapid, yielding transformant colonies in 48 to 72 h. Efficiencies of 10(sup5) to 10(sup6) transformants per (mu)g of plasmid DNA were reproducibly achieved for S. rimosus R6 and its mutants, and these numbers were 10(sup2) to 10(sup3) higher than those attained by polyethylene glycol-assisted transformation of protoplasts. In addition, we show that electroporation can be appli...Continue Reading

References

Apr 1, 1991·Applied and Environmental Microbiology·G M DunnyD J LeBlanc
Aug 15, 1990·Analytical Biochemistry·J M DiverP A Sokol
Oct 1, 1990·Journal of General Microbiology·S BonnassieA M Sicard
Aug 25, 1989·Nucleic Acids Research·D MattanovichH Katinger
Sep 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·S B SnapperW R Jacobs
Jul 11, 1988·Nucleic Acids Research·W J DowerC W Ragsdale
Feb 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·J F MillerL S Tompkins
Mar 1, 1970·Canadian Journal of Microbiology·V S Malik, L C Vining
Nov 1, 1982·Applied and Environmental Microbiology·J PigacM Alacević
Mar 1, 1988·Applied and Environmental Microbiology·Ian B PowellBarrie E Davidson
Mar 1, 1994·Applied and Environmental Microbiology·M SträtzP Dürre

❮ Previous
Next ❯

Citations

Jan 1, 1996·Critical Reviews in Biotechnology·S Y Ho, G S Mittal
Sep 9, 2006·Microbiology and Molecular Biology Reviews : MMBR·Hrvoje PetkovićPaul F Long
Jul 3, 2007·Molecular Plant-microbe Interactions : MPMI·Madhumita V Joshi, Rosemary Loria
Jan 31, 2009·Protein Engineering, Design & Selection : PEDS·Akbar NayeemJonathan Basch
Nov 6, 2010·Current Protocols in Microbiology·S Eric NyboJürgen Rohr
Mar 27, 2010·Letters in Applied Microbiology·S PhornphisutthimasA Thamchaipenet
Jul 31, 2016·Applied and Environmental Microbiology·Annika RöttigAlexander Steinbüchel
Oct 30, 2016·The Journal of Biological Chemistry·Jasna LalićIvan Ahel
Apr 3, 2016·Applied and Environmental Microbiology·Tina NetzkerAxel A Brakhage
Jun 9, 2007·Molecular Plant-microbe Interactions : MPMI·Madhumita JoshiRosemary Loria
Dec 1, 2009·Applied Microbiology and Biotechnology·Trond Erik Vee Aune, Finn Lillelund Aachmann
Apr 27, 2013·The Journal of Microbiology·Yong-Qiang FanYu-Long Wang
May 24, 2019·Applied Microbiology and Biotechnology·Jan KormanecLubomira Feckova
Jun 15, 1997·FEMS Microbiology Letters·C Mazy-ServaisJ Dusart
Feb 3, 2005·Molecular Microbiology·Johan A KersRosemary Loria
Oct 9, 2007·Molecular Microbiology·Madhumita V JoshiRosemary Loria
Apr 18, 1998·Applied and Environmental Microbiology·N B FitzgeraldT J Vanden Boom
Feb 9, 2018·Applied Microbiology and Biotechnology·Andrés Felipe Carrillo RincónHrvoje Petković
Aug 6, 2003·FEMS Microbiology Letters·Sofia StinchiMargherita Sosio
Feb 12, 2017·Applied Microbiology and Biotechnology·Yu DengXiaojuan Zhang
Oct 1, 1995·Applied and Environmental Microbiology·C PitulleG E Fox

❮ Previous
Next ❯

Related Feeds

Bacterial Cell Wall Structure (ASM)

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.

Bacterial Cell Wall Structure

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.