Expression in Neurospora
Dr W Dorsey Stuart
dorsey at UHUNIX.UHCC.HAWAII.EDU
Fri May 26 17:30:45 EST 1995
This may be of interest to academic labs thinking about expression in
> 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.
Dan Ebbole replied:
> 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
> 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.
The Neugenesis corporation of Honolulu Hawaii is in fact actively
developing expression systems in Neurospora. We are currently
collaborating with a major international corporation to produce a kit
which will contain expression vectors containing both constitutive and
regulated promoters and a cell line which has been engineered for
overexpression of recombinant proteins,
> currently grg-1 and qa-2 are the only regulated promoters that seem to have
> attracted attention for regulated overexpression of genes in Neurospora.
Our system utilizes three regulated promoters and two constitutive
promoters and we expect it to be available to academic labs this year.
As we develop the kit, we will be looking for a small number of labs to
become "alpha" testers of the system. If you are interested in trying
out a model version of our kit sometime within the next 60-90 days please
let me know.
Thanks and Aloha
W. Dorsey Stuart
Department of Genetics and Molecular Biology
School of Medicine
University of Hawaii
dorsey at uhunix.uhcc.hawaii.edu
Honolulu Hawaii 96822
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