Highly efficient gene targeting in the Aspergillus niger kusA mutant.
Gene targeting frequencies in Aspergillus niger are often very low and hamper efficient functional genomics in this biotechnologically important fungus. Deletion of the A. niger kusA gene encoding the ortholog of the Ku70 protein in other eukaryotes, dramatically improved homologous integration efficiency and reached more than 80% compared to 7% in the wild-type background, when 500bp homologous flanks were used. Furthermore, the use of the DeltakusA strain resulted in a high frequency of heterokaryon formation (70%) in primary transformants in the case disrupting an essential gene. Deletion of kusA had no obvious effect on the growth of the fungus, but renders the DeltakusA strain 10 times more sensitive to X-ray irradiation and two to three times more sensitive to UV exposure. The highly efficient gene targeting in combination with the A. niger genome sequence allows a systematic approach to generate gene knockouts and will help in improving the capacities of A. niger as producer of commercially interesting proteins and metabolites.
Meyer
Vera
V
Leiden University, Institute of Biology Leiden, Molecular Microbiology, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands.
Arentshorst
Mark
M
El-Ghezal
Aymen
A
Drews
Ann-Christin
AC
Kooistra
Rolf
R
van den Hondel
Cees A M J J
CA
Ram
Arthur F J
AF
D001234:Aspergillus niger / Q000235:genetics* / Q000254:growth & development / Q000528:radiation effects; D018390:Gene Targeting; D005800:Genes, Fungal / Q000235:genetics*; D009154:Mutation; D014466:Ultraviolet Rays; D014965:X-Rays