PMID- 17703973
OWN - NLM
STAT- MEDLINE
DCOM- 20080306
LR  - 20240104
IS  - 1096-0937 (Electronic)
IS  - 1087-1845 (Linking)
VI  - 45
IP  - 3
DP  - 2008 Mar
TI  - Transient disruption of non-homologous end-joining facilitates targeted genome 
      manipulations in the filamentous fungus Aspergillus nidulans.
PG  - 165-70
AB  - We have developed a transiently disrupted nkuA system in Aspergillus nidulans for 
      efficient gene targeting. The nkuA disruption was made by inserting a 
      counter-selectable marker flanked by a direct repeat (DR) composed of nkuA 
      sequences. In the disrupted state, the non-homologous end-joining (NHEJ) activity 
      is abolished and gene targeting can be performed with success rates identical to 
      those obtained with permanent nkuA knock-out strains. When gene targeting is 
      complete, the functional nkuA allele can be re-established via a simple selection 
      step, thereby eliminating the risk that defective NHEJ influences subsequent 
      analyses of the manipulated strain. Our system will facilitate construction of 
      large numbers of defined mutations in A. nidulans. Moreover, as the system can 
      likely be adapted to other filamentous fungi, we expect it will be particularly 
      beneficial in species where NHEJ cannot be restored by sexual crossing.
FAU - Nielsen, Jakob B
AU  - Nielsen JB
AD  - Center for Microbial Biotechnology, BioCentrum-DTU, Technical University of 
      Denmark, Building 223, DK-2800 Kgs. Lyngby, Denmark.
FAU - Nielsen, Michael L
AU  - Nielsen ML
FAU - Mortensen, Uffe H
AU  - Mortensen UH
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20070720
PL  - United States
TA  - Fungal Genet Biol
JT  - Fungal genetics and biology : FG & B
JID - 9607601
SB  - IM
MH  - Aspergillus nidulans/*genetics
MH  - Blotting, Southern
MH  - Fungi/*genetics
MH  - Gene Targeting/methods
MH  - Genes, Fungal/*genetics
MH  - Genetic Vectors/genetics
MH  - *Genome, Fungal
MH  - Mutation
MH  - Polymerase Chain Reaction
MH  - Recombination, Genetic
MH  - Transformation, Genetic
EDAT- 2007/08/21 09:00
MHDA- 2008/03/07 09:00
CRDT- 2007/08/21 09:00
PHST- 2007/05/24 00:00 [received]
PHST- 2007/06/15 00:00 [revised]
PHST- 2007/07/09 00:00 [accepted]
PHST- 2007/08/21 09:00 [pubmed]
PHST- 2008/03/07 09:00 [medline]
PHST- 2007/08/21 09:00 [entrez]
AID - S1087-1845(07)00120-X [pii]
AID - 10.1016/j.fgb.2007.07.003 [doi]
PST - ppublish
SO  - Fungal Genet Biol. 2008 Mar;45(3):165-70. doi: 10.1016/j.fgb.2007.07.003. Epub 
      2007 Jul 20.