Hi all!
Want to first of all send a collective thanks! to all those who sent
on a reply to my question - the rapidity of the response was
unbelievable and I got some good ideas. Again, thanks!
Secondly, for those who might be interested: was directed to a paper
in Mol. Cell. Biology (1987) 7 (3): 1180-1192 (Fleer et al.). in
which a yeast sequence which was unpropagatable in E. coli was
isolated from yeast. The cells are initially transformed into
spheroplasts and from here, an alkaline lysis was performed much like
that done with E. coli. CsCl gradients were performed and they were
able to recover 600-800ng of centromeric, 14kb plasmid from a liter of
minimal medium grown cells at a density of 8x10 7 cells per mL (out of
a total of 1.2ug isolated which included the 2mu plasmid - but if
working in cir- strains, contamination should be minimized, I would
think?
Hope this reference helps other interested parties. I know also of
another repair gene that was impossible to clone in E. coli, but what
it is escapes me for the moment.
Kathy
