Bifunctional protein conferring enhanced green fluorescence and puromycin resistance.
A new genetic marker was created in which sequences from enhanced green fluorescent protein were fused to those of puromycin N-acetyl transferase. The resulting fusion protein (EGFP-puro) conferred both green fluorescence and resistance to puromycin when expressed in mammalian cells. The utility of EGFP-puro as a selectable/screenable marker was demonstrated by the ease with which a recombinant guinea pig cytomegalovirus containing EGFP-puro was isolated by a combination of puromycin selection and screening for green fluorescence. We conclude that EGFP-puro is a compact and versatile marker that should prove useful for recombinant virus and transgenic cell line construction, particularly in applications in which coding capacity is limited.
Abbate
J
J
Governor's School for Government and International Studies, Richmond, VA, USA.
Lacayo
J C
JC
Prichard
M
M
Pari
G
G
McVoy
M A
MA
D000818:Animals; D000903:Antibiotics, Antineoplastic; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D003587:Cytomegalovirus / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D017930:Genes, Reporter; D005819:Genetic Markers; D049452:Green Fluorescent Proteins; D006168:Guinea Pigs; D006564:Herpesviridae / Q000235:genetics; D006801:Humans; D007202:Indicators and Reagents / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics*; D008168:Lung / Q000166:cytology; D016254:Mutagenesis, Insertional / Q000379:methods; D011691:Puromycin; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012867:Skin / Q000166:cytology
D000903:Antibiotics, Antineoplastic; D005819:Genetic Markers; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D011691:Puromycin