lambda-bank screening for proteases

Markus Woischnik MARKUS at scorpio.tng.oche.de
Fri May 26 10:13:00 EST 1995


Hi everybody,

I'm trying to "fish" protease-genes out of an lambda-bank made from  
genomic DNA with inserts from roughly 2-7kb, utilizing Stratagene's kit.
The phage infects E.coli Blue MRF and can be excised with an helper phage  
in order to clone a selected insert or the entire bank in E.coli XLOLR.

I can measure proteolytic activity in my culture-supernatant, but I don't  
have any sequence-information of the extracellular protease(s), so I can't  
use a probe for hybridisation-reactions.
On the other hand I don't want to purify and sequence the protein.

May be the proteases are expressed in their inactive and unexported form  
as preproprotein.

I'd appreciate if I can get any helpful hints !

thanks,
        markus
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