Going green in Cryptococcus neoformans: The recycling of a selectable drug marker |
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| Authors: | Raunak D Patel Jennifer K Lodge Lorina G Baker |
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| Institution: | 1. Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil;2. Departamento de Produtos Farmacêuticos, Faculdade de Farmácia, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil;3. Departamento de Patologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil;4. Laboratório de Micologia, Universidade Ceuma (UNICEUMA), São Luís, Maranhão, Brazil |
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| Abstract: | Cryptococcus neoformans is an opportunistic fungal pathogen that primarily affects immunocompromised individuals. Reverse genetics is commonly used to identify and characterize genes involved in a variety of cellular processes. In C. neoformans there is a limited set of positive selectable markers available to make gene deletions or other genetic manipulations. This has hampered the application of reverse genetics in this organism. We have adapted the Bacteriophage P1 Cre-loxP system for use in C. neoformans and successfully excised and reused the same drug marker, G418, to make two sequential gene deletions, lac1Δ and cap59Δ, in the same strain. This tool will allow investigators to make multiple sequential gene deletions in the same strain, which should facilitate the analysis of multigene families. |
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