Antibody-based strategy to identify Candida albicans genes expressed during infections.
Investigators have long used antibody-based screening strategies to identify Candida albicans immunogenic proteins and the genes that encode them during infections. With the recent availability of the C. albicans genome sequence and the development of genomic and proteomic technologies, it is now possible to efficiently conduct large-scale screening in standard research labs. C. albicans proteins and genes identified with a variety of screening methods have been implicated as important determinants of candidal virulence and exploited as vaccine and therapeutic targets. In this chapter, we describe methods used in our lab, in which sera recovered from patients with candidiasis are used to screen a C. albicans genomic DNA expression library. Immunoreactive colonies are detected by reaction with anti-human immunoglobulin, and the corresponding open reading frames are identified using the genome sequence database. The methods are also suitable for use with cDNA expression libraries, and they are complementary to proteomic screening strategies described elsewhere in this volume.
Clancy
Cornelius J
CJ
University of Florida College of Medicine and North Florida/South Georgia Veterans Health System, Gainesville, Florida, USA.
Cheng
Shaoji
S
Nguyen
M Hong
MH
D000908:Antibodies, Fungal; D000946:Antigens, Fungal / Q000235:genetics; D002176:Candida albicans / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D002177:Candidiasis / Q000276:immunology / Q000382:microbiology; D004271:DNA, Fungal / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005656:Fungal Proteins / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D015723:Gene Library; D005800:Genes, Fungal; D023281:Genomics; D054884:Host-Pathogen Interactions / Q000235:genetics / Q000276:immunology; D006801:Humans; D040901:Proteomics
D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D004271:DNA, Fungal; D005656:Fungal Proteins