A selectable bifunctional beta-galactosidase::phleomycin-resistance fusion protein as a potential marker for eukaryotic cells.
The Sh ble gene, conferring phleomycin resistance (PhR), was fused in frame to both the 3' and 5' ends of the Escherichia coli lacZ gene. The bifunctionality of the resulting 130-kDa hybrid proteins was demonstrated in E. coli and in the fungus, Tolypocladium geodes. PhR transformants of both organisms could be selected for. All transformants from E. coli and most from T. geodes displayed beta Gal activity. In the fungal host, higher transformation frequencies and greater levels of beta Gal activity were observed in clones harboring the lacZ::Sh ble fusion, as compared to the Sh ble::lacZ configuration. This system appears to be a potentially useful tool for the direct selection of transformants, and the evaluation of gene expression and regulation in a wide variety of prokaryotic and eukaryotic hosts.
Baron
M
M
Laboratoire de Microbiologie, Université Paul Sabatier, Toulouse, France.
Reynes
J P
JP
Stassi
D
D
Tiraby
G
G
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004351:Drug Resistance / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D005819:Genetic Markers; D003904:Mitosporic Fungi / Q000187:drug effects / Q000235:genetics; D008969:Molecular Sequence Data; D010692:Phleomycins / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics*
D004274:DNA, Recombinant; D005819:Genetic Markers; D010692:Phleomycins; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase