title: The use of direct cDNA selection to rapidly and effectively identify genes in the fungus Aspergillus fumigatus.
abstract: Aspergillus fumigatus is one of the causes of invasive lung disease in immunocompromised individuals. To rapidly identify genes in this fungus, including potential targets for chemotherapy, diagnostics, and vaccine development, we constructed cDNA libraries. We began with non-normalized libraries, then to improve this approach we constructed a normalized cDNA library using direct cDNA selection. Normalization resulted in a reduction of the frequency of clones with highly expressed genes and an enrichment of underrepresented cDNAs. Expressed sequence tags generated from both the original and the normalized libraries were compared with the genomes of Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Candida albicans, indicating that a large proportion of A. fumigatus genes do not have orthologs in these fungal species. This method allowed the expeditious identification of genes in a fungal pathogen. The same approach can be applied to other human or plant pathogens to rapidly identify genes without the need for genomic sequence information.
authors.affiliation: Genome Therapeutics Corporation, Waltham, MA 02453, USA. marco.kessler@genomecorp.com
authors.forename: Marco M
authors.initials: MM
authors.lastname: Kessler
authors.forename: Debra Aker
authors.initials: DA
authors.lastname: Willins
authors.forename: Qiandong
authors.initials: Q
authors.lastname: Zeng
authors.forename: Richard G
authors.initials: RG
authors.lastname: Del Mastro
authors.forename: Robin
authors.initials: R
authors.lastname: Cook
authors.forename: Lynn
authors.initials: L
authors.lastname: Doucette-Stamm
authors.forename: Hongmei
authors.initials: H
authors.lastname: Lee
authors.forename: Anne
authors.initials: A
authors.lastname: Caron
authors.forename: Terri K
authors.initials: TK
authors.lastname: McClanahan
authors.forename: Luquan
authors.initials: L
authors.lastname: Wang
authors.forename: Jonathan
authors.initials: J
authors.lastname: Greene
authors.forename: Roberta S
authors.initials: RS
authors.lastname: Hare
authors.forename: Guillaume
authors.initials: G
authors.lastname: Cottarel
authors.forename: George H
authors.initials: GH
authors.lastname: Shimer
mesh_terms: D001232:Aspergillus fumigatus / Q000235:genetics*; D001483:Base Sequence; D002176:Candida albicans / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics*; D020224:Expressed Sequence Tags; D015723:Gene Library; D005800:Genes, Fungal; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D004271:DNA, Fungal