ISSN:
1432-0983
Keywords:
Phleomycin resistance
;
Transformation
;
Fungi
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary A convenient and efficient transformation system has been developed for the filamentous fungus Tolypocladium geodes. In contrast to most of the commonly described techniques requiring prior preparation of protoplasts or spheroplasts, this method leads to high efficiency transformation of T. geodes conidiospores following moderate lytic enzyme treatment. Competent cells so obtained are still resistant to osmotic pressure and can be stored frozen without loss of viability. The highest transformation frequency (3-5x103 transformants per μg of DNA) was obtained with plasmid pUT737 containing the Sh ble gene conferring phleomycin resistance under the control of a strong promoter isolated from Trichoderma reesei. Southern hybridization revealed multiple integration sites of plasmid DNA into the T. geodes nuclear DNA despite the absence of homology between the transforming DNA and the recipient genome. Instability could not be detected for the phleomycin® phenotype during more than five generations of mitotic growth under non-selective conditions.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00318520
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