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Paxilline-negative mutants of Penicillium paxilli generated by heterologous and homologous plasmid integration.

Authors :
Young, Carolyn
Itoh, Yasuo
Johnson, R.
Garthwaite, I.
Miles, Christopher O.
Munday-Finch, Sarah C.
Scott, B.
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