Penicillium Transformation & Selection

Stephan Martin Summerer smsummer at acs.ucalgary.ca
Thu Aug 31 17:39:54 EST 1995


Hello Ted,
	I have been doing enough transformations of P. urticae to
feell confident in recomending phleomycin (or it's sister
compound bleomycin). It's expensive but you don't need much
(~15ug/mL) for efficient selection. For a protocol I use the
standard protoplasts and PEG (I tried electroporation and didn't
have any luck at all). When you begin to optimize a protocol, one
variable that you should look at is the transforming DNA. I found
that linear DNA gave a >~2 fold increase and that methylation
state of DNA also made a difference (but for me it became more of
a bother to isolated unmethylated DNA for E. coli GM2163 than
just using double the amount of TG2 derived plasmid DNA)
	Write me if you need any more info.
	Stephan

In article <424ent$228 at bigboote.WPI.EDU>,
Theodore C. Crusberg <crusberg at wpi.edu> wrote:
>I am working with Penicillium ochro-chloron and would like to be able
>to transform this sp.  First, we need a selectable marker, but the
>organism is already hygromycin B resistant.  Any ideas?  We also need
>a transformation protocol for Penicillium (protoplasts
>vs. electroporation vs. balistic methods?), and even a vector
>(plasmid?)  We currently have no auxotrophs and would prefer some kind
>of antibiotic selectable marker. Please help if you can.
>Ted Crusberg/Biology & Biotechnology Dept./Worcester Polytechnic
>Institute/Worcester MA 01609
>





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