The use of transposons as molecular tools for random mutagenesis in filamentous fungi

Ref ID: 18483

Author:

Linda Paun, ElKbir Hihlal, Frank Kempken

Author address:

Abteilung für Botanische Genetik und Molekularbiologie, Botanisches Institut, Christian-Albrechts-Universität zu
Kiel

Full conference title:

11 th European Conference on Fungal Genetics

Abstract:

The mobile and mostly also repetitive genetic elements named transposons are found in all eukaryotic genomes.
Their impact on gene expression as well as their ability to cause major mutations within the genome or single
genes makes them an interesting tool for random mutagenesis. Using transposable elements in filamentous fungi
would aid gene characterization for instance in pathogenic or biotechnological important strains (1, 2, 3). At
current we are employing two types of transposons, i.e. Restless from Tolypocladium inflatum (4) and Vader from
Aspergillus niger (5). Each transposable element was inserted between the promoter and the open reading frame
of the hygromycin phosphotransferase (hph) gene and transformed into Neurospora crassa or Aspergilli,
respectively. We use hygromycin selection to identify transposition events and observed a Vader excision
frequency of about 1 in 2.2 x 10
5
spores. All colonies analyzed showed an excision event on the DNA level. Vader
footprints were found and after performing TAIL8208;PCRs the reintegration sites of 21 independent excision events
were determined. Vader mostly integrates within or very close to genes, thus it appears to be a useful tool for
transposon8208;mediated mutagenesis in A. niger (6). At current we try to improve the Vader tool and analyze its
function in the heterologous host A. nidulans. Furthermore we analyse the performance of Restless in N. crassa.  Â 
1. Kempken F, Kück U (1998) BioEssays 20:6528208;659
2. Kempken F (2003) In: Arora DK, Khachatourians GG (eds) Applied Mycology and Biotechnology, Vol. 3 Fungal
Genomics, Elsevier Science Annual Review Series, pp838208;99
3.Pöggeler S, Kempken F (2004) In: Kück U (ed) The Mycota II, Genetics and Biotechnology, 2nd edition, Springer
Verlag, Heidelberg, New York, Tokyo, pp1658208;198
4.Kempken F, Kück U (1996) Mol Cell Biol 16:65638208;6572
5. Braumann I, van den Berg M, & Kempken F (2007) Fungal Genet Biol 44(12):13998208;1414.  
6. Hihlal E, Braumann I, van den Berg M, Kempken F (2011) Appl Environment Microbiol, 77: 2332-2336

Abstract Number: PR3.32

Conference Year: 2012

Link to conference website: http://www.ecfg.info/images/Abstract_Book_Electronic.pdf

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