Transient expression of the beta-glucuronidase gene after biolistic transformation of the anaerobic fungus Neocallimastix frontalis.
The rumen anaerobic fungus Neocallimastix frontalis was biolistically transformed using plasmids containing the bacterial beta-glucuronidase gene (GUS) fused to the promoter sequences of the enolase gene from N. frontalis. Multiple copies of the plasmids were precipitated onto tungsten particles and delivered into zoosporangia and a mycelial mat by a helium-driven biolistic device. Transformants were detected by histochemical assay for beta-glucuronidase. It was found that the enolase promoter sequences tested were responsible for the transient expression of the beta-glucuronidase gene. This is the first study presenting results on the transformation of an anaerobic fungus.
Durand
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R
Laboratoire de Biologie Cellulaire Fongique, Centre de Génétique Moléculaire et Cellulaire, CNRS UMR 5534, Bât 405, Université Claude Bernard Lyon I, F-69622 Villeurbanne Cedex, France.
Rascle
C
C
Fischer
M
M
Fèvre
M
M
D000693:Anaerobiosis; D000818:Animals; D019470:Biolistics; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D005658:Fungi / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D005798:Genes, Bacterial; D005966:Glucuronidase / Q000235:genetics* / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000235:genetics; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012417:Rumen / Q000382:microbiology; D013172:Spores, Fungal / Q000235:genetics; D014170:Transformation, Genetic
D004271:DNA, Fungal; D005966:Glucuronidase; D010751:Phosphopyruvate Hydratase