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Applied and Environmental Microbiology, November 2000, p. 4810-4816, Vol. 66, No. 11
Department of
Microbiology1 and Institute of Legal
Medicine,2 Medical School of the University
of Innsbruck, A-6020 Innsbruck, Austria
Received 5 June 2000/Accepted 17 August 2000
A highly inducible fungal promoter derived from the
Penicillium chrysogenum endoxylanase (xylP)
gene is described. Northern analysis and the use of a
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Copyright © 2000, American Society for Microbiology. All rights reserved.
xylP Promoter-Based Expression System and Its Use for
Antisense Downregulation of the Penicillium chrysogenum
Nitrogen Regulator NRE
-glucuronidase (uidA) reporter gene strategy showed that
xylP expression is transcriptionally regulated. Xylan and
xylose are efficient inducers, whereas glucose strongly represses the
promoter activity. Comparison of the same expression construct as a
single copy at the niaD locus in P. chrysogenum and at the argB locus in Aspergillus nidulans
demonstrated that the xylP promoter is regulated similarly
in these two species but that the level of expression is about 80 times
higher in the Aspergillus species. The xylP
promoter was found to be 65-fold more efficient than the
isopenicillin-N-synthetase (pcbC) promoter in
Penicillium and 23-fold more efficient than the nitrate
reductase (niaD) promoter in Aspergillus under
induced conditions. Furthermore, the xylP promoter was used
for controllable antisense RNA synthesis of the nre-encoded
putative major nitrogen regulator of P. chrysogenum. This
approach led to inducible downregulation of the steady-state mRNA level
of nre and consequently to transcriptional repression of
the genes responsible for nitrate assimilation. In addition, transcription of nreB, which encodes a negative-acting
nitrogen regulatory GATA factor of Penicillium, was found
to be subject to regulation by NRE. Our data are the first direct
evidence that nre indeed encodes an activator in the
nitrogen regulatory circuit in Penicillium and
indicate that cross regulation of the controlling factors occurs.
*
Corresponding author. Mailing address: Department of
Microbiology (Medical School), University of Innsbruck,
Fritz-Pregl-Str. 3, A-6020 Innsbruck, Austria. Phone: 43-512-507-3608. Fax: 43-512-507-2866. E-mail:
hubertus.haas{at}uibk.ac.at.
Applied and Environmental Microbiology, November 2000, p. 4810-4816, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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