1887

Abstract

The gene encoding yeast-enhanced green fluorescent protein (GFP) was placed under control of ALS gene promoters in . The P-GFP reporter strains were validated using various techniques including a new real-time RT-PCR assay to quantify ALS gene expression. The P-GFP reporter strains were grown in media that promoted yeast or germ tube forms, and the resulting fluorescence was measured by flow cytometry. In addition to results that indicate differences in ALS gene expression due to growth medium, growth stage and developmental programme, new data show large differences in transcriptional level among the ALS genes. Expression of was associated with transfer of the P-GFP strain to fresh growth medium. expression increased markedly when germ tubes were visible microscopically and expression exhibited a transient peak between 2 and 3 h following inoculation into fresh YPD medium. Transcription from the and promoters was strongest among those tested and contrasted markedly with the weaker promoter strength at the , , and loci. These weaker transcriptional responses were also observed using real-time RT-PCR measurements on wild-type cells. Assuming a positive correlation between transcriptional level and protein production, these results suggest that some Als proteins are abundant on the cell surface while others are produced at a much lower level.

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2005-04-01
2019-11-19
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