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Paxilline-negative mutants of Penicillium paxilli generated by heterologous and homologous plasmid integration.
- Source :
- Current Genetics; Jun1998, Vol. 33 Issue 5, p368-377, 10p
- Publication Year :
- 1998
-
Abstract
- Using a monoclonal antibody based ELISA, 600 pAN7-1 plasmid-tagged mutants of Penicillium paxilli were screened for paxilline accumulation and one paxilline-negative mutant, YI-20, was identified. A molecular analysis of this mutant showed that pAN7-1 was inserted at a single site but was present as 4–6 copies arranged in a head-to-tail tandem array. Rescue of flanking sequences and analysis of the corresponding genomic region revealed that YI-20 has an extensive deletion at the site of pAN7-1 integration. Probing of a CHEF gel with the same sequences showed that associated with the deletion is a rearrangement of chromosome Va. Targeted gene disruption of wild-type sequences adjacent to the site where pAN7-1 inserted, resulted in the generation of two additional paxilline-negative mutants; both were single crossovers with deletions extending outside the region mapped. Neither of these new mutants had a rearrangement of chromosome Va, suggesting that deletion of genes on this chromosome is responsible for the paxilline-negative phenotype. Telomeric fingerprinting of genomic digests of P. paxilli, combined with pulsed-field gel electrophoresis of chromosomal DNA, established that there are a minimum of eight chromosomes in this fungus. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 01728083
- Volume :
- 33
- Issue :
- 5
- Database :
- Complementary Index
- Journal :
- Current Genetics
- Publication Type :
- Academic Journal
- Accession number :
- 49957645
- Full Text :
- https://doi.org/10.1007/s002940050349