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New pFA‐cassettes for PCR‐based gene manipulation in Candida albicans
Author(s) -
Schaub Yvonne,
Dünkler Alexander,
Walther Andrea,
Wendland Jürgen
Publication year - 2006
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.200510133
Subject(s) - candida albicans , marker gene , gene , auxotrophy , corpus albicans , biology , transformation (genetics) , computational biology , microbiology and biotechnology , selectable marker , chemistry , genetics , escherichia coli
Abstract Several modules for efficient PCR‐based gene disruption have recently been introduced in Candida albicans . These are based on auxotrophic marker genes for deficient strains derived from SC5314/CAI4. Commonly used protocols for the transformation C. albicans are based either on the lithium acetate procedure or on electroporation also used for Saccharomyces cerevisiae . Here we present our updated arsenal of pFA‐modules that now include the heterologous marker genes HIS1 from C. dubliniensis and LEU2 from C. maltosa ( Noble and Johnson 2005) and the dominant selection marker ca SAT1 (Reuss et al. 2004). We also introduce the Ashbya gossypii TEF1 ‐promoter as a strong constitutive promoter. With these new elements an enlarged collection of pFA‐marker and pFA‐marker‐promoter modules were generated containing 17 new modules. In addition, N‐terminal tagging with GFP‐(GA) 6 and epitope‐tagging modules using the 6 ×‐HIS‐tag were constructed. This adds to the previous modules that only enabled C‐terminal GFP‐tagging of genes ( Gola et al. 2003). In total 29 pFA‐modules are currently freely available from our lab which – together with an update on the diagnostic verification procedure – further enlarge the C. albicans molecular toolbox and enhance our capabilities to use PCR‐based gene alteration methods in C. albicans . (© 2006 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)