Abstract：
:Aspergillus nidulans is an important experimental organism, and it is a model organism for the genus Aspergillus that includes serious pathogens as well as commercially important organisms. Gene targeting by homologous recombination during transformation is possible in A. nidulans, but the frequency of correct gene targeting is variable and often low. We have identified the A. nidulans homolog (nkuA) of the human KU70 gene that is essential for nonhomologous end joining of DNA in double-strand break repair. Deletion of nkuA (nkuA delta) greatly reduces the frequency of nonhomologous integration of transforming DNA fragments, leading to dramatically improved gene targeting. We have also developed heterologous markers that are selectable in A. nidulans but do not direct integration at any site in the A. nidulans genome. In combination, nkuA delta and the heterologous selectable markers make up a very efficient gene-targeting system. In experiments involving scores of genes, 90% or more of the transformants carried a single insertion of the transforming DNA at the correct site. The system works with linear and circular transforming molecules and it works for tagging genes with fluorescent moieties, replacing genes, and replacing promoters. This system is efficient enough to make genomewide gene-targeting projects feasible.
journal_name
Geneticsjournal_title
Geneticsauthors
Nayak T,Szewczyk E,Oakley CE,Osmani A,Ukil L,Murray SL,Hynes MJ,Osmani SA,Oakley BRdoi
10.1534/genetics.105.052563keywords:
subject
Has Abstractpub_date
2006-03-01 00:00:00pages
1557-66issue
3eissn
0016-6731issn
1943-2631pii
genetics.105.052563journal_volume
172pub_type
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