Bacillus transformation

Frank Skraly skralyf at cae.wisc.edu
Mon Nov 27 13:22:54 EST 1995


I need to express heterologous proteins in Bacillus (e.g. subtilis).

My first attempt, with an E. coli/gram-positive shuttle vector with Km 
resistance, failed.  I couldn't even get any transformants.  I tried 
another (Cm-resistance) plasmid and got transformants, but that plasmid
had already been cut and pasted beyond usefulness, so I couldn't do much
with it.  

Does anyone have a suggestion on what vector you've used in Bacillus that
enabled you to (inducibly, even) express heterologous proteins and have 
decent stability?  The copy number does not have to be high.  I also don't
want the proteins to be secreted.

Thank you for any and all suggestions!

-----
Frank Skraly
University of Wisconsin-Madison
skralyf at cae.wisc.edu



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