In article <4b3jh7$fnd at trst.cca.vu.nl>, jos urbanus <urbanus at chem.vu.nl> wrote:
> Hi, everybody
> I am from the biochemical department of the VU, Amsterdam, and trying to
> develop different kinds of toxicity assays with yeast. In order to
> increase the availability of a drug to a cell, spheroplasts can play an
> important role in combination with certain kind of drugs. In such an
> assay it would be essential that most cells stay alive and keep on
> proliferating.
> To produce spheroplasts from normal cells is very easy, but until now I
> have not been able to regenerate these cells. In an 0.7 M sorbitol
> YNB-medium spheroplast do not generate a cell wall, produce enormous
> vacuoles and finally blow themselves up, I guess.
> So, if anybody has experience with spheroplast regeneration, please give
> me advice or a protocol.
Try 1 M sorbitol.
--
Michael Lichten
lichten at helix.nih.gov
