Going green in Cryptococcus neoformans: the recycling of a selectable drug marker.

Autor: Patel RD; Department of Molecular Microbiology, Washington University School of Medicine, 660 S. Euclid Avenue, Saint Louis, MO 63110, USA., Lodge JK, Baker LG
Jazyk: angličtina
Zdroj: Fungal genetics and biology : FG & B [Fungal Genet Biol] 2010 Mar; Vol. 47 (3), pp. 191-8. Date of Electronic Publication: 2009 Nov 26.
DOI: 10.1016/j.fgb.2009.11.007
Abstrakt: Cryptococcus neoformans is an opportunistic fungal pathogen that primarily affects immunocompromised individuals. Reverse genetics is commonly used to identify and characterize genes involved in a variety of cellular processes. In C. neoformans there is a limited set of positive selectable markers available to make gene deletions or other genetic manipulations. This has hampered the application of reverse genetics in this organism. We have adapted the Bacteriophage P1 Cre-loxP system for use in C. neoformans and successfully excised and reused the same drug marker, G418, to make two sequential gene deletions, lac1Delta and cap59Delta, in the same strain. This tool will allow investigators to make multiple sequential gene deletions in the same strain, which should facilitate the analysis of multigene families.
(Copyright (c) 2009 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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