Nuclear labeling of Coccidioides posadasii with green fluorescent protein

Lei Li, Monika Schmelz, Ellen M. Kellner, John N. Galgiani, Marc J. Orbach

Research output: Chapter in Book/Report/Conference proceedingConference contribution

6 Scopus citations

Abstract

Coccidioidomycosis is a mild to life-threatening disease in otherwise healthy humans and other mammals caused by the fungus Coccidioides spp. Understanding the development of the unique dimorphic life cycle of Coccidioides spp. and its role in pathogenesis has been an area of research focus. However, nuclear behavior during the saprobic and parasitic life cycle has not been studied intensively. In this study, green fluorescent protein (GFP)was fused to histone H1 and introduced into Coccidioides posadasii (C. posadasii) strain Silveira to monitor the nuclear behavior of the fungus during the saprobic and parasitic stages of the life cycle. We constructed an Agrobacterium tumefaciens-mediated transformation (ATMT) vector that had in its T-DNA region a hygromycin-resistance gene aswell as the fused histone H1-GFP gene under the control of the histone H3 promoter of C. posadasii. More than 30 hygromycin-resistant transformants were obtained and 23 were purified to homozygosity throughmultiple passages of the original transformants on hygromycin-containing media. One strain (VFC1420) transformed with a single copy of the fusion histone H1-GFP gene was selected for cytological studies. Strong nuclear-localized GFP signals were observed in arthroconidia, hyphae, as well as in spherules and endospores developed in vitro. Thus GFP can be used to study the expression pattern of potential virulence genes identified in serial analysis of gene expression (SAGE) or expressed sequence tags (EST) libraries, and could be a useful tool to monitor disease development in the murine model.

Original languageEnglish (US)
Title of host publicationCoccidioidomycosis Sixth International Symposium
PublisherBlackwell Publishing Inc.
Pages198-207
Number of pages10
ISBN (Print)1573316881, 9781573316880
DOIs
StatePublished - Sep 2007

Publication series

NameAnnals of the New York Academy of Sciences
Volume1111
ISSN (Print)0077-8923
ISSN (Electronic)1749-6632

Keywords

  • Coccidioides
  • Coccidioidomycosis
  • GFP
  • Green fluorescent protein
  • Valley Fever

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • History and Philosophy of Science

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    Li, L., Schmelz, M., Kellner, E. M., Galgiani, J. N., & Orbach, M. J. (2007). Nuclear labeling of Coccidioides posadasii with green fluorescent protein. In Coccidioidomycosis Sixth International Symposium (pp. 198-207). (Annals of the New York Academy of Sciences; Vol. 1111). Blackwell Publishing Inc.. https://doi.org/10.1196/annals.1406.014