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• W657290304 abstract "This chapter presents the strategy developed to generate a Candida albicans ORFeome collection in a versatile Gateway vector, which allows the transfer of the cloned genes into a variety of C. albicans Gateway-compatible expression vectors. The development of collections of over-expression strains is facilitated by the pre-establishment of a complete set of cloned open reading frames (ORFs), or ORFeome. Importantly, ORFeomes represent useful resources for the implementation of other approaches used to elucidate gene function apart from gene deletion or overexpression. Individual characterization of mutants on the genome-wide scale can be slow and laborious. In this respect, signature-tagged mutagenesis (STM) provides an attractive alternative. In STM, each mutant is tagged with a different DNA sequence, allowing all tags to be amplified from the DNA of mixed populations of mutants in a single PCR. In the C. albicans ORFeome, the start codon of the 6,205 ORFs has been included, whereas the stop codon has been excluded to allow insertion of C-terminal tags. The authors have favored C-terminal tagging since tags at the amino termini may possibly interfere with targeting of proteins to the secretory pathway. A panel of libraries of strains expressing tagged proteins should facilitate the systematic execution of high-throughput biochemical and microscopic assays of the C. albicans proteome. The construction of the ORFeome could pave the way to the generation of the C. albicans promoterome (library of gene promoters) in order to better characterize expression profiles." @default.
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• W657290304 date "2014-04-09" @default.
• W657290304 modified "2023-09-26" @default.
• W657290304 title "Cool Tools 5: The Candida albicans ORFeome Project" @default.
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• W657290304 doi "https://doi.org/10.1128/9781555817176.ch34" @default.
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