[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.
Goldenkova
I V
IV
Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, 119991 Russia. irengold@vigg.ru
Musiĭchuk
K A
KA
Piruzian
E S
ES
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