By Erika J. Ernst
A suite of cutting-edge molecular equipment for learning antifungal resistance, for locating and comparing either new and current antifungal medicinal drugs, and for figuring out the host reaction and immunotherapy of such brokers. The protocols stick with the profitable equipment in Molecular drugs™ sequence structure, every one providing step by step laboratory directions, an advent outlining the main in the back of the strategy, lists of the mandatory apparatus and reagents, and pointers on troubleshooting and keeping off recognized pitfalls. Antifungal brokers: tools and Protocols deals clinician-scientists, microbiologists and molecular biologists the efficient instruments they want this day to appreciate and effectively boost new healing brokers for yeast, mould, and fungal infections.
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Extra resources for Antifungal Agents: Methods And Protocols (Methods in Molecular Medicine)
Albicans mutants, however, the MPAR marker has to be recycled after insertion into the first allele of the target gene to allow its subsequent use in a second round of transformation for inactivation of the remaining wild-type allele. Because a negative selection against the presence of the MPAR marker is not available in C. albicans wild-type strains, we developed a method that allowed the specific excision of the marker from the genome of transformants with high efficiency. We designed a cassette, the MPAR flipper, which in addition to the MPAR marker, contains a C.
4. , and Morschhäuser, J. (2000) Targeted gene disruption in Candida albicans wild-type strains: the role of the MDR1 gene in fluconazole resistance of clinical Candida albicans isolates. Mol. Microbiol. 36, 856–865. 5. Köhler, G. , White, T. , and Agabian, N. (1997) Overexpression of a cloned IMP dehydrogenase gene of Candida albicans confers resistance to the specific inhibitor mycophenolic acid. J. Bacteriol. 179, 2331–2338. 6. , et al. (1999) Host-induced, stage-specific virulence gene activation in Candida albicans during infection.
Remove solution C with a 10-mL pipet, cover the gel with 20X SSC, and transfer the DNA onto the nylon membrane by vacuum blotting for 90 min. 9. 4 N NaOH for 30 s. 10. 5, for 30 s. 11. Place the membrane on a sheet of Whatman paper to remove excess liquid and fix the DNA on the nylon membrane using a UV crosslinker. 32 Morschhäuser, Staib, and Köhler 12. Rinse the membrane briefly in 5X SSC, put it in a hybridization oven bottle (DNA side should face the interior, not the glass walls), and add 15 mL of ECL hybridization solution prewarmed to 42°C.