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New method enables gene disruption in destructive fungal pathogen

Researchers at the Virginia Bioinformatics Institute (VBI) at Virginia Tech, Colorado State University, and Duke University Medical Center have developed a new method to determine gene function on a genome-wide scale in the fungal pathogen Alternaria brassicicola. This destructive fungus causes black spot disease, leading to considerable leaf loss in such economically important crops as canola, cabbage, and broccoli.

Genomic methods that allow the disruption of several thousand genes are needed because they allow high-throughput identification of genes and gene function. Such procedures are widely applicable and would be extremely useful in allowing scientists to investigate the key events that occur when a host interacts with a pathogen.

"The development of this protocol is timely as the genome sequence of A. brassicicola is scheduled for completion in 2006. We now have in our hands a versatile method that will allow us to dissect the pathogen's nucleotide sequence information and establish the function of many of the individual genes in this filamentous fungus," said Christopher Lawrence, associate professor at VBI, director of the project, and one of the authors of the study.

"A. brassicicola has consistently been used in studies with the weedy mustard plant Arabidopsis. The genome sequence of Arabidopsis was determined in 2001 and many methods are available to ascertain gene function in this plant," Lawrence said. "We now have a means to identify key fungal and plant genes that interact and ultimately lead to disease development or resistance. This is an extremely powerful research tool."

The generation of gene disruption mutants has been a limiting step for the analysis of gene function in most filamentous fungi. The new method takes advantage of a novel linear DNA construct that greatly improves the efficiency of targeted gene disruption. The DNA construct includes an antibiotic-resistance marker gene, which allows for easy sel
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Source:Virginia Tech


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