By Erika J. Ernst
A set of state of the art molecular equipment for learning antifungal resistance, for locating and comparing either new and present antifungal medications, and for figuring out the host reaction and immunotherapy of such brokers. The protocols stick to the winning equipment in Molecular medication™ sequence structure, each one delivering step by step laboratory directions, an creation outlining the main in the back of the approach, lists of the mandatory apparatus and reagents, and pointers on troubleshooting and fending off identified pitfalls. Antifungal brokers: equipment and Protocols deals clinician-scientists, microbiologists and molecular biologists the efficient instruments they wish this present day to appreciate and effectively enhance new healing brokers for yeast, mould, and fungal infections.
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Extra info for Antifungal Agents
Albicans). If no correct clones are obtained after screening of a reasonable number of MPA-resistant transformants (approx 20–30 clones) it is better to repeat the transformation with a freshly prepared disruption cassette. 3. Transformation of C. albicans can cause undesired genomic alterations. To make sure that phenotypes are the result of inactivation of the target gene, construct at least two independent homozygous mutants, starting from two independent first round transformants. In addition, an intact copy of the target gene should be reintegrated into the homozygous mutants, and this should revert the mutant phenotype.
Targeted Gene Deletion in C. albicans Fig. 1. 39 40 40 Morschhäuser, Staib, and Köhler Fig. 2. Selection of mycophenolic acid (MPA)-resistant Candida albicans transformants and screening for MPA-sensitive derivatives in which the MPAR flipper cassette was excised by FLP-mediated recombination. (A) A C. albicans wild-type strain was transformed with a deletion construct containing the MPAR flipper cassette inserted between flanking sequences of a target gene. The photograph was taken after 7 d of growth of the transformants at 30°C on an synthetic defined (SD) agar plate containing 10 µg/ mL MPA.
40, 2300–2305. 3. , and Staib, P. (1999) Sequential gene disruption in Candida albicans by FLP-mediated site-specific recombination. Mol Microbiol 32, 547–556. 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.