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A simple medium for rapid regeneration of Bacillus subtilis protoplasts transformed with plasmid DNA

1987; Oxford University Press; Volume: 40; Issue: 1 Linguagem: Inglês

10.1111/j.1574-6968.1987.tb01972.x

1574-6968

Antonio Puyet, H. Sandoval, Paloma LÃ pez, Alfredo Aguilar, José Martín, Manuel Espinosa,

Bacterial Genetics and Biotechnology

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.