Transcriptional activation by recombinant GAL4-VP16 in the Xenopus oocyte

In addition to measuring transcription in vitro or in transient transfection assays, an alternative method to study the function of transcription factors is to inject reporter DNA together with either nuclear extract or factor gene into Xenopus oocytes. Here we introduce the GAL4-derived assay, whic...

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Bibliographic Details
Published in:Nucleic acids research 1993-06, Vol.21 (11), p.2775-2775
Main Authors: Xu, Licen, Schaffner, Walter, Rungger, Duri
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Bivalvia
Cell Nucleus - metabolism
DNA - metabolism
DNA-Binding Proteins
Female
Fundamental and applied biological sciences. Psychology
Fungal Proteins - metabolism
Gene Expression
Globins - biosynthesis
Herpes Simplex Virus Protein Vmw65 - metabolism
Molecular and cellular biology
Molecular genetics
Nuclear Proteins - genetics
Oocytes - metabolism
Recombinant Fusion Proteins - metabolism
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins
Transcription Factors - metabolism
Transcription, Genetic
Transcription. Transcription factor. Splicing. Rna processing
Xenopus
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Summary:In addition to measuring transcription in vitro or in transient transfection assays, an alternative method to study the function of transcription factors is to inject reporter DNA together with either nuclear extract or factor gene into Xenopus oocytes. Here we introduce the GAL4-derived assay, which was established in other systems, to study transcriptional activation in the frog oocyte. In our experiment, purified recombinant transactivator fusion protein GAL-VP16(40N) was coinjected with a beta -globin reporter gene into the Xenopus oocytes nucleus (germinal vesicle). The reporter promoter contained five GAL4 binding sites upstream of an octamer motif. Xenopus oocytes were prepared as described previously.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/21.11.2775