Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/68633
標題: Agrobacterium T-DNA-mediated integration and gene replacement in the brown rot pathogen Monilinia fructicola
作者: Lee, M.H.
Bostock, R.M.
關鍵字: cutinase
T-DNA tandem integration
nucleotide deletion
filler
multinucleate
fungi
saccharomyces-cerevisiae genome
neurospora-crassa
aspergillus-awamori
illegitimate recombination
insertional mutagenesis
functional
genomics
botrytis-cinerea
hygromycin-b
plant-cells
transformation
期刊/報告no:: Current Genetics, Volume 49, Issue 5, Page(s) 309-322.
摘要: A transformation system utilizing Agrobacterium tumefaciens was developed for targeted gene disruption in Monilinia fructicola, a fungal pathogen that causes brown rot disease of stone fruits. Transformation with a vector containing the neomycin phosphotransferase II (nptII) cassette flanked with 4 kb cutinase gene (Mfcut1) flanking sequences resulted in an average of 13 transformants per 10(5) spores. When assayed by PCR and DNA blot analyses, more than 50% of the transformants recovered had integrated in the targeted Mfcut1 locus. Both target-gene-specific and non-specific integrations carried direct (head-to-tail) repeat T-DNA integrations. Sequence analysis of these T-DNA integrations revealed that 26 bp of the T-DNA right border were missing at the junctions between direct repeats in all cases. The recombination event during non-specific T-DNA integration in this fungus was unlike that reported in Agrobacterium-mediated transformation in plants.
URI: http://hdl.handle.net/11455/68633
ISSN: 0172-8083
文章連結: http://dx.doi.org/10.1007/s00294-006-0059-0
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