Expression in Neurospora

Dan Ebbole dje0282 at VMS2.TAMU.EDU
Fri May 26 13:22:16 EST 1995


Geoff Turner
 University of Sheffield  found:

We wish to overexpress N. crassa glutamate dehydrogenase, 
since we are working on its structure, and E. coli is 
found to give insoluble, inactive protein.
We turned to N. crassa, and used the grg-1 expression vector 
deposited at the FGSC.  Our initial transformants with 
grg-1 driving the am gene (=gdh) are rather disappointing.

Interesting observation.  I've not noticed that subculturing leads to a
loss of expression of genes in transformed strains. Our experience has been
that different levels of expression can be obtained with SOME promoters
even when integrated at the same position in the genome.  I'm not sure how
stable this expression is during subculturing.  With the con-10 gene
expression is reproducible between independent transformants and over time
with culturing.  So it looks like there may be some Grg-1 specific problem.
 I guess these are ectopic transformants and unsure how this might affect
things.  

I would indeed consider expression in A. nidulans since the expression
systems are better worked out for Aspergillus species.   I do think that
Neurospora is a "suitable" host given some effort, but you (and I, and
everybody else) have time constraints that prevent a real effort to develop
expression systems in Neurospora when they might not be any better than
those already developed for Aspergillus.  

currently grg-1 and qa-2 are the only regulated promoters that seem to have
attracted attention for regulated overexpression of genes in Neurospora. 
------------------------
Dr. Daniel Ebbole
dje0282 at summa.tamu.edu
Assistant Professor
Dept. of Plant Pathology and Microbiology
Texas A&M University
College Station, TX 77843-2132
USA




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