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PMID: 7476854 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Tagging pathogenicity genes in Ustilago maydis by restriction enzyme-mediated integration (REMI).

Molecular & general genetics : MGG ·Vol. 248 ·No. 5 ·1995-09-20 ·Pages 547-52

Bölker M, Böhnert HU, Braun KH, Görl J, Kahmann R

Abstract

In the maize pathogenic fungus Ustilago maydis integration of transforming DNA at homologous or heterologous sites is often accompanied by duplications of the DNA. We show that it is possible to generate single-copy integration events with high efficiency by restriction enzyme-mediated integration (REMI). In about 50% of cases, a plasmid that contains a single BamHI site is integrated at chromosomal BamH1 sites, if BamHI is added to the transformation mixtures. In the other cases it appears that integration events have also occurred preferentially at BamHI sites, but without restoration of the recognition sites. Using REMI we have generated approximately 1000 insertion mutants. Pathogenicity tests demonstrated that about 1-2% of these mutants were unable to induce symptoms when tested in planta. For two of the mutants we have shown that the phenotype is linked to the insertion event.

MeSH Terms
Base Sequence DNA Restriction Enzymes Molecular Sequence Data Mutagenesis, Insertional Plants/microbiology Plasmids Restriction Mapping Ustilago/genetics,pathogenicity
Chemicals
DNA Restriction Enzymes
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Bölker M
Institut für Genetik und Mikrobiologie, Universität München, Germany.
Böhnert H U
Braun K H
Görl J
Kahmann R
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21 references, click to expand
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF02423450
Published
1995-09-20
Pages
547-52
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF02423450
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