1887

Abstract

An RT-PCR assay was developed to analyse expression patterns of genes in the ALS (gglutinin-ike equence) family. Inoculation of a reconstituted human buccal epithelium (RHE) model of mucocutaneous candidiasis with strain SC5314 showed destruction of the epithelial layer by and also formation of an upper fungal layer that had characteristics similar to a biofilm. RT-PCR analysis of total RNA samples extracted from -inoculated buccal RHE showed that , , , , and were consistently detected over time as destruction of the RHE progressed. Detection of transcripts from , and particularly from , was more sporadic, but not associated with a strictly temporal pattern. The expression pattern of ALS genes in cultures used to inoculate the RHE was similar to that observed in the RHE model, suggesting that contact of with buccal RHE does little to alter ALS gene expression. RT-PCR analysis of RNA samples extracted from model denture and catheter biofilms showed similar gene expression patterns to the buccal RHE specimens. Results from the RT-PCR analysis of biofilm RNA specimens were consistent between various strains during biofilm development and were comparable to gene expression patterns in planktonic cells. The RT-PCR assay described here will be useful for analysis of human clinical specimens and samples from other disease models. The method will provide further insight into the role of ALS genes and their encoded proteins in the diverse interactions between and its host.

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2004-02-01
2019-11-20
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