| Autor: |
Adachi, Kiichi, Nelson, Grant H., Peoples, Keith A., Frank, Sheryl A., Montenegro-Chamorro, Maria V., DeZwaan, Todd M., Ramamurthy, Lakshman, Shuster, Jeffrey R., Hamer, Lisbeth, Tanzer, Matthew M. |
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| Zdroj: |
Current Genetics; Aug2002, Vol. 42 Issue 2, p123-127, 5p |
| Abstrakt: |
TAGKO (transposon-arrayed gene knockout) is a highly efficient method for gene discovery and gene function assignment in the rice blast fungus Magnaporthe grisea. Here, we report the application of genome-wide TAGKO to the wheat blotch fungus Mycosphaerella graminicola, including the successful development of electroporation-based transformation for this fungus. A M. graminicola genomic cosmid library was constructed and a pool of 250 cosmid clones was mutagenized by in vitro transposition. Sequence analysis identified 5,110 unique insertion events in the M. graminicola genome. Eleven transposon-tagged cosmid clones (TAGKO clones) were chosen and transformed into the wild-type strain by electroporation. Ten TAGKO clones out of 11 produced gene-specific mutants at a targeting frequency of 15–28%, significantly higher than that of conventional gene-disruption constructs. The remaining clone failed to produce viable mutants, thereby providing indirect evidence for the identification of an essential gene. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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