Penicillium Transformation & Selection

Stephan Martin Summerer smsummer at
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.

In article <424ent$228 at bigboote.WPI.EDU>,
Theodore C. Crusberg <crusberg at> 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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