Development of a genetic transformation system for Histoplasma capsulatum: complementation of uracil auxotrophy |
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Authors: | Patricia L Worsham and William E Goldman |
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Institution: | (1) Department of Molecular Microbiology, Washington University School of Medicine, 63110 St. Louis, Mo, USA;(2) Present address: U.S. Army Medical Research Institute of Infectious Diseases, Frederick, Md, USA |
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Abstract: | Summary We have developed a simple and efficient transformation system for the dimorphic fungus Histoplasma capsulatum. Mutants of H. capsulatum defective in orotidine-5 -monophosphate pyrophosphorylase were transformed to prototrophy by the cloned URA5 gene of the filamentous fungus Podospora anserina. Abortive and mitotically stable transformants were obtained. The stable transformants had integrated copies of the plasmid, some in tandem head-to-tail orientation. Free plasmid identical to the transforming plasmid was present in some of the transformants. We obtained a transformation efficiency of up to 30 transformants/ g DNA for plasmid pPAura5-1 (9.2 kb). pPW2001, a smaller plasmid (4.7 kb) derived from pPAura5-1, transformed H. capsulatum more efficiently (up to 155 transformants/gm DNA). |
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Keywords: | Histoplasma capsulatum Fungus Transformation Uracil Auxotroph |
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