Open AccessA simple medium for rapid regeneration of Bacillus subtilis protoplasts transformed with plasmid DNA
Author(s) -
Puyet A.,
Sandoval H.,
López P.,
Aguilar A.,
Martin J.F.,
Espinosa M.
Publication year - 1987
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1987.tb01972.x
Subject(s) - protoplast , bacillus subtilis , plasmid , polyethylene glycol , bacteria , microbiology and biotechnology , sucrose , incubation , biology , bacillaceae , bacillales , dna , chemistry , biochemistry , genetics
A sucrose‐based simple medium (c‐R5) is described on which protoplasts prepared from Bacillus subtilis are able to revert rapidly to bacillary forms. Transformations of B. subtilis protoplasts were performed using polyethylene glycol and various plasmid DNAs, followed by regeneration of the protoplasts on c‐R5 medium. Regenerated bacteria were easily detectable on selective c‐R5 medium after only 16 h of incubation at 37°C. This medium can be used for direct selection of various antibiotic‐resistance markers. In addition, the c‐R5 medium can be simplified to make it chemically defined, allowing the selection of prototrophs.