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  • Title: Cyclosporin A-resistance based gene placement system for Neurospora crassa.
    Author: Bardiya N, Shiu PK.
    Journal: Fungal Genet Biol; 2007 May; 44(5):307-14. PubMed ID: 17320431.
    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.
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