Efficient gene identification and targeted gene disruption in the wheat blotch fungus Mycosphaerella graminicola using TAGKO

被引：15

作者：

Adachi, K ^{[1
]}

Nelson, GH ^{[1
]}

Peoples, KA ^{[1
]}

Frank, SA ^{[1
]}

Montenegro-Chamorro, MV ^{[1
]}

DeZwaan, TM ^{[1
]}

Ramamurthy, L ^{[1
]}

Shuster, JR ^{[1
]}

Hamer, L ^{[1
]}

Tanzer, MM ^{[1
]}

机构：

[1] Paradigm Genet Inc, Res Triangle Pk, NC 27709 USA

来源：

CURRENT GENETICS | 2002年 / 42卷 / 02期

关键词：

Mycosphaerella graminicola; Septoria wheat blotch; TAGKO; gene disruption; electroporation;

D O I：

10.1007/s00294-002-0339-2

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

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.

引用

页码：123 / 127

页数：5

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