Graduated resistance to G418 leads to differential selection of cultured mammalian cells expressing the neo gene.
The effects of Geneticin (G418) selection on the growth and survival of cultured mammalian cells expressing the neomycin-resistance gene (neo) were studied by the analysis of cell clones from two retroviral neo vector-infected populations. We found a correlation between the neo expression level and growth rates in medium containing varying G418 concentrations. This relationship permits the use of differential selection schemes for the isolation of rare cells with increased expression. Comparison, by clone sampling, of vector-positive populations before and after selection with a G418 concentration in the range usually used for selection, showed different expression level and vector copy number distributions for the population infected with the vector of lower LTR activity, but not for the other. Such biasing effects of G418 selection may be important when selected cells are used for quantitative studies of gene expression.
Paludan
K
K
Department of Molecular Biology and Plant Physiology, University of Aarhus, Denmark.
Duch
M
M
Jørgensen
P
P
Kjeldgaard
N O
NO
Pedersen
F S
FS
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002999:Clone Cells; D003001:Cloning, Molecular; D004351:Drug Resistance / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D005839:Gentamicins / Q000494:pharmacology*; D019868:Kanamycin Kinase; D007700:Kinetics; D009355:Neomycin / Q000494:pharmacology*; D010770:Phosphotransferases / Q000235:genetics* / Q000378:metabolism
D000900:Anti-Bacterial Agents; D005839:Gentamicins; C010680:antibiotic G 418; D010770:Phosphotransferases; D019868:Kanamycin Kinase; D009355:Neomycin