Fusion of green fluorescent protein with the Zeocin-resistance marker allows visual screening and drug selection of transfected eukaryotic cells.
Green fluorescent protein (GFP) and the Zeocin-resistance gene Sh ble (ZeoR) were fused together to generate a bifunctional protein for the identification and selection of transfected mammalian cells. Expression of this hybrid selectable marker, GFP-ZeoR, was visually detected and conferred Zeocin resistance in prokaryotes and eukaryotes. This selectable marker provides a way to determine transient transfection efficiencies in tissue culture cells using fluorescence microscopy. Expression of the GFP-ZeoR was also used to identify and select stable mammalian cell lines expressing a heterologous gene. Selection was efficient and GFP fluorescence provides an excellent, noninvasive technique to monitor the success of Zeocin selection during the development of the stable cell lines. This hybrid resistance gene combines the functional properties of the Zeocin-resistance marker and GFP and should be useful for combined selection and fluorescence in a variety of organisms.
Bennett
R P
RP
Invitrogen Corporation, Carlsbad, CA, USA.
Cox
C A
CA
Hoeffler
J P
JP
D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D001761:Bleomycin / Q000737:chemistry*; D016466:CHO Cells; D019556:COS Cells; D002460:Cell Line; D006224:Cricetinae; D003587:Cytomegalovirus / Q000235:genetics; D004351:Drug Resistance; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins; D000882:Haplorhini; D006801:Humans; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D010957:Plasmids / Q000096:biosynthesis / Q000138:chemical synthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics*; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection / Q000235:genetics*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C099353:phleomycin resistance protein, Streptoalloteichus hindustanus; D001761:Bleomycin; D049452:Green Fluorescent Proteins; C105427:Zeocin; D001616:beta-Galactosidase