Green fluorescent protein as a reporter for gene expression in the mucoralean fungus Absidia glauca

Schilde, C Wostemeyer, J Burmester, A

C. Schilde et al., Green fluorescent protein as a reporter for gene expression in the mucoralean fungus Absidia glauca, ARCH MICROB, 175(1), 2001, pp. 1-7

33

INGLESE

Article

Microbiology

ARCHIVES OF MICROBIOLOGY

0302-8933 → ACNP

175

1

2001

1 - 7

ISI

0302-8933(200101)175:1<1:GFPAAR>2.0.ZU;2-U

Mucoralean fungi (Zygomycota) are used for many industrial processes and al so as important model organisms for investigating basic biological problems . Their genetic analysis is severely hampered by low transformation frequen cies, by their strong tendency towards autonomous replication of plasmids i nstead of stable integration, and by the lack of reliable genetic reporter systems. We constructed plasmids for transforming the model zygomycete Absi dia glauca that carry the versatile reporter gene coding for green fluoresc ent protein (GFP). gfp expression is controlled either by the homologous ac tin promoter or the promoter for the elongation factor of translation, EF1 alpha. These plasmids also confer neomycin resistance and carry one of two genetic elements (rag1, seg1) that improve mitotic stability of the plasmid . The Rfp constructs were replicated extrachromosomally and could be recove red from retransformed Escherichia coli cells. gfp expression was monitored by epifluorescence microscopy. The gfp reporter gene plasmids presented he re for the model zygomycete A. glauca constitute the first reliable system that allows the monitoring of gene expression in this important group of fu ngi.