Bifunctional reporter genes: construction and expression in prokaryotic and eukaryotic cells |
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Authors: | Goldenkova I V Musiĭchuk K A Piruzian E S |
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Affiliation: | Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, 119991 Russia. irengold@vigg.ru |
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Abstract: | Bifunctional reporter proteins were constructed to combine Clostridium thermocellum lichenase (LicBM2) with Aequorea victoria green fluorescent protein (GFP) or with Escherichia coli beta-glucuronidase (GUS). The major properties of the initial proteins were preserved in the hybrid ones: LicBM2 was active at 65 degrees C, GFP fluoresced, and GUS hydrolyzed its substrates. LicBM2 remained active after extension of its C of N end. Bifunctional reporter systems were shown to provide a convenient tool for studying the gene expression regulation in prokaryotic (E. coli) and eukaryotic (Saccharomyces cerevisiae, mammalian) cells, advantages of one reporter compensating for drawbacks of the other. |
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