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PMID: 2381417 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Development of a genetic transformation system for Histoplasma capsulatum: complementation of uracil auxotrophy.

Molecular & general genetics : MGG ·Vol. 221 ·No. 3 ·1990-05-00 ·Pages 358-62

Worsham PL, Goldman WE

Abstract

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/micrograms 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/micrograms DNA).

MeSH Terms
DNA Replication DNA, Fungal/genetics Genetic Complementation Test Histoplasma/enzymology,genetics Mitosis Molecular Weight Orotate Phosphoribosyltransferase/metabolism Plasmids Transformation, Genetic Uracil
Chemicals
DNA, Fungal Uracil Orotate Phosphoribosyltransferase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Worsham P L
Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Mo 63110.
Goldman W E
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24 references, click to expand
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00259400
Published
1990-05-00
Pages
358-62
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
Grants
NIAID NIH HHS · AI25584 · United States
External Links
PubMed source DOI: 10.1007/BF00259400
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