title: Cyclosporin A-resistance based gene placement system for Neurospora crassa.
abstract: DNA introduced into Neurospora crassa are usually inserted at random ectopic sites of the genome, often in multiple copies. To facilitate the study of gene expression and function, transformation by a single-copy of a gene at a defined locus is desired. Although several targeted gene placement methods are available for N. crassa, they all require a specific genetic background in the recipient. We describe here the development of a new locus for targeted gene placement that does not require any pre-existing marker in the target strain. Our system takes advantage of the fact that disruption of the csr-1 gene, which encodes the cyclosporin A-binding protein, leads to the resistance to cyclosporin A. By cloning a gene of interest into a csr-1 knock-in vector and transforming a fungus with it, one can easily insert any gene, in single-copy, into a defined locus.
authors.affiliation: Division of Biological Sciences, University of Missouri, Columbia, MO 65211, USA.
authors.forename: Nirmala
authors.initials: N
authors.lastname: Bardiya
authors.forename: Patrick K T
authors.initials: PK
authors.lastname: Shiu
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology; D016572:Cyclosporine / Q000494:pharmacology*; D004271:DNA, Fungal; D025141:Drug Resistance, Fungal; D005800:Genes, Fungal; D005822:Genetic Vectors / Q000235:genetics; D008957:Models, Genetic; D009492:Neurospora crassa / Q000187:drug effects* / Q000235:genetics*; D014170:Transformation, Genetic
chemical_list: D000935:Antifungal Agents; D004271:DNA, Fungal; D016572:Cyclosporine