Fungal infection remains a major problem worldwide, yet treatment options are limited owing to the lack of effective drugs, the significant toxicity of available compounds, and the emergence of drug resistance. The low toxicity of calcofluor white(CFW) is an attractive antifungal compound for its known inhibitive effects on trichophyton rubrum and candida albicans growth. However, the efficacy of CFW is limited in most cases. In order to search for effective means to improve its efficacy, using saccharomyces cerevisiae as a model, we have used microarrays to examine the cell's response when treated with CFW on the genome scale. We found that both the PKC-SLT2(i.e, protein kinase C-mitogen activated protein kinase) and the glycogen metabolic pathways are activated upon CFW treatment. These results suggest that the key components in these pathways could be targeted by other drugs to counter the cell's compensative response, thus to further substantiate the inhibitive effect of CFW on fungal growth, which may lead to treatment regimens with improved efficacy of this compound in clinical applications.
With the accomplishment of the genome draft sequences, identification of functional elements in genome has become an urgent task. Full-length cDNAs provide an important resource for gene identification and their precise structural feature determination. It also provides a basis for genomic element definition. As many regulatory elements are around transcription start sites(TSSs), precise localization of TSSs in the genome becomes a critical step for identifying the associated core promoters. Massive parallel snapshot of TSSs at a particular time under a specific experimental condition makes it possible to globally analyze important regulatory elements around TSSs and further construct transcriptional regulatory networks. In this paper, we first reviewed two important full-length cDNA cloning techniques: cap-trapper technique and oligo-capping technique. Then,we introduced deepCAGE, a cap-trapper and deep sequencing-based TSS profiling technique, and its applications in the research of transcriptional regulation.
