[Bifunctional reporter genes: construction and expression in prokaryotic and eukaryotic cells].
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.
Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, 119991 Russia. irengold@vigg.ru
I V
IV
Goldenkova
K A
KA
Musiĭchuk
E S
ES
Piruzian
D000818:Animals; D003013:Clostridium / Q000235:genetics; D003063:Cnidaria / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005057:Eukaryotic Cells / Q000502:physiology*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008322:Mammals; D016716:PC12 Cells; D011387:Prokaryotic Cells / Q000502:physiology*; D015202:Protein Engineering / Q000379:methods*; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics
D008164:Luminescent Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D006026:Glycoside Hydrolases; C028020:licheninase