High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA.
A highly efficient method for transformation of Bacillus subtilis by plasmid DNA is reported. The procedure, which involves polyethylene glycol-induced DNA uptake by protoplasts and subsequent regeneration of the bacterial cell wall, yields up to 80% transformants with an efficiency of 4 x 10(7) transformants per microgram of supercoiled DNA. Plasmids constructed by in vitro ligation or endonuclease-generated fragments of linear plasmid DNA can also transform PEG-treated protoplasts, but at a lower frequency.
D001412:Bacillus subtilis / Q000235:genetics*; D002701:Chloramphenicol / Q000494:pharmacology; D004270:DNA, Circular / Q000235:genetics*; D007612:Kanamycin / Q000494:pharmacology; D007700:Kinetics; D011523:Protoplasts / Q000378:metabolism; D011815:R Factors; D014169:Transformation, Bacterial
D004270:DNA, Circular; D007612:Kanamycin; D002701:Chloramphenicol