Bifunctional protein conferring enhanced green fluorescence and puromycin resistance

被引:25
作者
Abbate, J
Lacayo, JC
Prichard, M
Pari, G
McVoy, MA
机构
[1] Virginia Commonwealth Univ, Dept Pediat, Richmond, VA 23298 USA
[2] Govenors Sch Govt & Int Studies, Richmond, VA USA
[3] Hybridon, Milford, MA USA
关键词
D O I
10.2144/01312st05
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:336 / +
页数:4
相关论文
共 25 条
[1]   THE PROKARYOTIC NEOMYCIN-RESISTANCE-ENCODING GENE ACTS AS A TRANSCRIPTIONAL SILENCER IN EUKARYOTIC CELLS [J].
ARTELT, P ;
GRANNEMANN, R ;
STOCKING, C ;
FRIEL, J ;
BARTSCH, J ;
HAUSER, H .
GENE, 1991, 99 (02) :249-254
[2]   A SELECTABLE BIFUNCTIONAL BETA-GALACTOSIDASE--PHLEOMYCIN-RESISTANCE FUSION PROTEIN AS A POTENTIAL MARKER FOR EUKARYOTIC CELLS [J].
BARON, M ;
REYNES, JP ;
STASSI, D ;
TIRABY, G .
GENE, 1992, 114 (02) :239-243
[3]  
Baum C, 1996, EXP HEMATOL, V24, P364
[4]   Fusion of green fluorescent protein with the Zeocin™-resistance marker allows visual screening and drug selection of transfected eukaryotic cells [J].
Bennett, RP ;
Cox, CA ;
Hoeffler, JP .
BIOTECHNIQUES, 1998, 24 (03) :478-482
[5]   The selectable marker neo gene down-regulates gene expression from retroviral vectors containing an internal ribosome entry site [J].
Byun, J ;
Kim, JM ;
Robbins, PD ;
Kim, S .
GENE THERAPY, 1998, 5 (10) :1441-1444
[6]   gpt-gus fusion gene for selection and marker in recombinant poxviruses [J].
Cao, JX ;
Upton, C .
BIOTECHNIQUES, 1997, 22 (02) :276-278
[7]   A BIFUNCTIONAL FUSION BETWEEN BETA-GLUCURONIDASE AND NEOMYCIN PHOSPHOTRANSFERASE - A BROAD-SPECTRUM MARKER ENZYME FOR PLANTS [J].
DATLA, RSS ;
HAMMERLINDL, JK ;
PELCHER, LE ;
CROSBY, WL ;
SELVARAJ, G .
GENE, 1991, 101 (02) :239-246
[8]   POSITIVE SELECTABLE MARKERS FOR USE WITH MAMMALIAN-CELLS IN CULTURE [J].
EGLITIS, MA .
HUMAN GENE THERAPY, 1991, 2 (03) :195-201
[9]   QUANTITATIVE-ANALYSIS OF GENE SUPPRESSION IN INTEGRATED RETROVIRUS VECTORS [J].
EMERMAN, M ;
TEMIN, HM .
MOLECULAR AND CELLULAR BIOLOGY, 1986, 6 (03) :792-800
[10]   PROMOTER TRAPS IN EMBRYONIC STEM-CELLS - A GENETIC SCREEN TO IDENTIFY AND MUTATE DEVELOPMENTAL GENES IN MICE [J].
FRIEDRICH, G ;
SORIANO, P .
GENES & DEVELOPMENT, 1991, 5 (09) :1513-1523