Deletion of the Trichodiene Synthase Gene of Fusarium venenatum:Two Systems for Repeated Gene Deletions

Autor: Royer, John C, Christianson, Lynne M, Yoder, Wendy T, Gambetta, Greg A, Klotz, Alan V, Morris, Carin L, Brody, Howard, Otani, Suzie
Zdroj: Fungal Genetics and Biology; October 1999, Vol. 28 Issue: 1 p68-78, 11p
Abstrakt: The trichodiene synthase (tri5) gene of Fusarium venenatumwas cloned from a genomic library. Vectors were created in which the tri5coding sequence was replaced with the Neurospora crassanitrate reductase (nit3) gene and with the Aspergillus nidulansacetamidase (amdS) gene flanked by direct repeats. The first vector was utilized to transform a nitrate reductase (niaD) mutant of F. venenatumto prototrophy, and the second vector was utilized to confer acetamide utilization to the wild-type strain. Several of the transformants lost the capacity to produce the trichothecene diacetoxyscirpenol and were shown by hybridization analysis to have gene replacements at the tri5locus. The nit3gene was removed by retransformation with a tri5deletion fragment and selection on chlorate. The amdSgene was shown to excise spontaneously via the flanking direct repeats when spores were plated onto fluoroacetamide.
Databáze: Supplemental Index
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