- 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 & Permissions
- CITING ARTICLES
- Citing Articles via HighWire
- Citing Articles via Google Scholar
- GOOGLE SCHOLAR
- Articles by Steiner, S.
- Articles by Philippsen, P.
- Search for Related Content
- PUBMED
- PubMed Citation
- Articles by Steiner, S.
- Articles by Philippsen, P.
Genetics, Vol 140, 973-987, Copyright © 1995
INVESTIGATIONS |
Homologous Recombination as the Main Mechanism for DNA Integration and Cause of Rearrangements in the Filamentous Ascomycete Ashbya gossypii
S. Steiner, J. Wendland, M. C. Wright and P. Philippsen
Institut fur Mikrobiologie und Molekularbiologie, Justus-Liebig-Universitat, Giessen, D-35392 Giessen, Germany and Institut fur Angewandte Mikrobiologie, Biozentrum Basel, CH-4056 Basel, Switzerland.
A slow and a fast growth phenotype were observed after transformation of the phytopathogenic fungus Ashbya gossypii using a plasmid carrying homologous DNA and as selectable marker the Tn903 aminoglycoside resistance gene expressed from a strong A. gossypii promoter. Transformations with circular plasmids yielded slowly and irregularly growing geneticin-resistant mycelia in which 1% of nuclei contained plasmid sequences. Occasionally, fast growing sectors appeared which were shown to be initiated by homologous integration of the transforming DNA. Transformants obtained with plasmids linearized within the homology region immediately exhibited fast radial growth. In all 28 transformants analyzed plasmid DNA was integrated homologously. Such apparent lack of nonhomologous recombination has so far not been observed in filamentous ascomycetes. In 14 transformants two to four tandemly integrated plasmid copies were found. They underwent several types of genetic changes, mainly in the older mycelium: excision of whole plasmid copies and rearrangements within the integrated DNA (inversions and deletions). These internal rearrangements involved 360-bp inverted repeats, remnants of IS-elements flanking the resistance gene, and 156-bp direct repeats, originating from the strong A. gossypii promoter. Improved vectors lacking sequence repetitions were constructed and used for stable one-step gene replacement in A. gossypii.
This article has been cited by other articles:
 |
|
 |
|
  P. Knechtle, J. Wendland, and P. Philippsen The SH3/PH Domain Protein AgBoi1/2 Collaborates with the Rho-Type GTPase AgRho3 To Prevent Nonpolar Growth at Hyphal Tips of Ashbya gossypii Eukaryot. Cell, October 1, 2006; 5(10): 1635 - 1647. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H.-P. Schmitz, A. Kaufmann, M. Kohli, P. P. Laissue, and P. Philippsen From Function to Shape: A Novel Role of a Formin in Morphogenesis of the Fungus Ashbya gossypii Mol. Biol. Cell, January 1, 2006; 17(1): 130 - 145. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Jimenez, M. A. Santos, M. Pompejus, and J. L. Revuelta Metabolic Engineering of the Purine Pathway for Riboflavin Production in Ashbya gossypii Appl. Envir. Microbiol., October 1, 2005; 71(10): 5743 - 5751. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Hermida, S. Brachat, S. Voegeli, P. Philippsen, and M. Primig The Ashbya Genome Database (AGD)--a tool for the yeast community and genome biologists Nucleic Acids Res., January 1, 2005; 33(suppl_1): D348 - D352. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. S. Dietrich, S. Voegeli, S. Brachat, A. Lerch, K. Gates, S. Steiner, C. Mohr, R. Pohlmann, P. Luedi, S. Choi, et al. The Ashbya gossypii Genome as a Tool for Mapping the Ancient Saccharomyces cerevisiae Genome Science, April 9, 2004; 304(5668): 304 - 307. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Knechtle, F. Dietrich, and P. Philippsen Maximal Polar Growth Potential Depends on the Polarisome Component AgSpa2 in the Filamentous Fungus Ashbya gossypii Mol. Biol. Cell, October 1, 2003; 14(10): 4140 - 4154. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Schlosser, G. Schmidt, and K.-P. Stahmann Transcriptional regulation of 3,4-dihydroxy-2-butanone 4-phosphate synthase Microbiology, December 1, 2001; 147(12): 3377 - 3386. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Wendland and P. Philippsen Cell Polarity and Hyphal Morphogenesis Are Controlled by Multiple Rho-Protein Modules in the Filamentous Ascomycete Ashbya gossypii Genetics, February 1, 2001; 157(2): 601 - 610. [Abstract] [Full Text]
|
|
|
|
 |
|
 C Alberti-Segui, F Dietrich, R Altmann-Johl, D Hoepfner, and P Philippsen Cytoplasmic dynein is required to oppose the force that moves nuclei towards the hyphal tip in the filamentous ascomycete Ashbya gossypii J. Cell Sci., January 3, 2001; 114(5): 975 - 986. [Abstract] [PDF]
|
|
|
|
|
|
Y Ayad-Durieux, P Knechtle, S Goff, F Dietrich, and P Philippsen A PAK-like protein kinase is required for maturation of young hyphae and septation in the filamentous ascomycete Ashbya gossypii J. Cell Sci., January 12, 2000; 113(24): 4563 - 4575. [Abstract] [PDF]
|
|
|
|
|
|
J Wendland and P Philippsen Determination of cell polarity in germinated spores and hyphal tips of the filamentous ascomycete Ashbya gossypii requires a rhoGAP homolog J. Cell Sci., January 5, 2000; 113(9): 1611 - 1621. [Abstract] [PDF]
|
|
|
|
 |
|
 C. Forster, M. A. Santos, S. Ruffert, R. Kramer, and J. L. Revuelta Physiological Consequence of Disruption of the VMA1 Gene in the Riboflavin Overproducer Ashbya gossypii J. Biol. Chem., April 2, 1999; 274(14): 9442 - 9448. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Monschau, H. Sahm, and K.-P. Stahmann Threonine Aldolase Overexpression plus Threonine Supplementation Enhanced Riboflavin Production in Ashbya gossypii Appl. Envir. Microbiol., November 1, 1998; 64(11): 4283 - 4290. [Abstract] [Full Text]
|
|