This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Brakhage, A. A. Articles by Van den Brulle, J. Search for Related Content PubMed PubMed Citation Articles by Brakhage, A. A. Articles by Van den Brulle, J.

 Previous Article  |  Next Article 

J. Bacteriol., 05 1995, 2781-2788, Vol 177, No. 10
Copyright © 1995, American Society for Microbiology

Use of reporter genes to identify recessive trans-acting mutations specifically involved in the regulation of Aspergillus nidulans penicillin biosynthesis genes

AA Brakhage and J Van den Brulle
Lehrstuhl fur Mikrobiologie, Universitat Munchen, Federal Republic of Germany.

Starting from three amino acid precursors, penicillin biosynthesis is catalyzed by three enzymes which are encoded by the following three genes: acvA (pcbAB), ipnA (pcbC), and aat (penDE). To identify trans- acting mutations which are specifically involved in the regulation of these secondary metabolism genes, a molecular approach was employed by using an Aspergillus nidulans strain (AXTII9) carrying acvA-uidA and ipnA-lacZ gene fusions integrated in double copies at the chromosomal argB gene. On minimal agar plates supplemented with X-Gal (5-bromo-4- chloro-3-indolyl-beta-D-galactopyranoside), colonies of such a strain stained blue, which is indicative of ipnA-lacZ expression. After mutagenesis with UV light, colonies were isolated on agar plates with lactose as the carbon source, which produced only a faint blue color or no color at all. Such mutants (named Prg for penicillin regulation) most likely were defective in trans-acting genes. Control experiments revealed that the mutants studied still carried the correct number of gene fusions. In a fermentation run, mutants Prg-1 and Prg-6 exhibited only 20 to 50% of the ipnA-lacZ expression of the wild-type strain and produced only 20 to 30% of the penicillin produced by the wild-type strain. Western blot (immunoblot) analysis showed that these mutants contained reduced amounts of ipnA gene product, i.e., isopenicillin N synthase. Both mutant Prg-1 and mutant Prg-6 also differed in acvA-uidA expression levels from the wild type. Segregation analysis indicated that for both mutants the Prg phenotype resulted from mutation of a single gene.(ABSTRACT TRUNCATED AT 250 WORDS)

This article has been cited by other articles:

• Grosse, C., Heinekamp, T., Kniemeyer, O., Gehrke, A., Brakhage, A. A. (2008). Protein Kinase A Regulates Growth, Sporulation, and Pigment Formation in Aspergillus fumigatus. Appl. Environ. Microbiol. 74: 4923-4933 [Abstract] [Full Text]  
• Thon, M., Al-Abdallah, Q., Hortschansky, P., Brakhage, A. A. (2007). The Thioredoxin System of the Filamentous Fungus Aspergillus nidulans: IMPACT ON DEVELOPMENT AND OXIDATIVE STRESS RESPONSE. J. Biol. Chem. 282: 27259-27269 [Abstract] [Full Text]  
• Mander, G. J., Wang, H., Bodie, E., Wagner, J., Vienken, K., Vinuesa, C., Foster, C., Leeder, A. C., Allen, G., Hamill, V., Janssen, G. G., Dunn-Coleman, N., Karos, M., Lemaire, H. G., Subkowski, T., Bollschweiler, C., Turner, G., Nusslein, B., Fischer, R. (2006). Use of laccase as a novel, versatile reporter system in filamentous fungi.. Appl. Environ. Microbiol. 72: 5020-5026 [Abstract] [Full Text]  
• Herrmann, M., Sprote, P., Brakhage, A. A. (2006). Protein Kinase C (PkcA) of Aspergillus nidulans Is Involved in Penicillin Production. Appl. Environ. Microbiol. 72: 2957-2970 [Abstract] [Full Text]  
• Liebmann, B., Muller, M., Braun, A., Brakhage, A. A. (2004). The Cyclic AMP-Dependent Protein Kinase A Network Regulates Development and Virulence in Aspergillus fumigatus. Infect. Immun. 72: 5193-5203 [Abstract] [Full Text]  
• Langfelder, K., Philippe, B., Jahn, B., Latge, J.-P., Brakhage, A. A. (2001). Differential Expression of the Aspergillus fumigatus pksP Gene Detected In Vitro and In Vivo with Green Fluorescent Protein. Infect. Immun. 69: 6411-6418 [Abstract] [Full Text]  
• Martín, J. F. (2000). Molecular Control of Expression of Penicillin Biosynthesis Genes in Fungi: Regulatory Proteins Interact with a Bidirectional Promoter Region. J. Bacteriol. 182: 2355-2362 [Full Text]  
• Van den Brulle, J., Steidl, S., Brakhage, A. A. (1999). Cloning and Characterization of an Aspergillus nidulans Gene Involved in the Regulation of Penicillin Biosynthesis. Appl. Environ. Microbiol. 65: 5222-5228 [Abstract] [Full Text]  
• Brakhage, A. A. (1998). Molecular Regulation of beta -Lactam Biosynthesis in Filamentous Fungi. Microbiol. Mol. Biol. Rev. 62: 547-585 [Abstract] [Full Text]