TY - JOUR
T1 - Puromycin resistance in Chinese hamster cells
T2 - Genetic and biochemical studies of partially resistant, unstable clones
AU - Morrow, John
AU - Sammons, David
AU - Barron, Emily
N1 - Funding Information:
1300 York Ave, New York, NY 10012 (U.S.A.) This work was supported by a grant from the Public Health Service (1-R01-CA-1607-01).
PY - 1980/2
Y1 - 1980/2
N2 - Resistance to 10 μg/ml of puromycin has been analyzed in V79 Chinese hamster cells. Clones that were isolated in 10 μg/ml of puromycin and subsequently cultivated in its absence consistently lost their resistance. One clone was analyzed in detail by recloning in the presence and absence of puromycin, and it was found that non-puromycin cultivated sublones also lost their resistance and regained inhibition profiles similar to the V79 parent. Reconstruction experiments between sensitive and resistant cells demonstrated that the yield of mutants was not affected by metabolic cooperation. The mutation rate was calculated to be 1 × 10-7 per cell per generation, and was the same within the limits of statistical error in a colchicine-produced polyploid derivative of the V79 line. Although a number of resistant clones were found to have polyploid karyotypes, the polyploid V79 line was not more resistant to puromycin, nor did it possess a higher frequency of puromycin resistant cells. Studies employing radiolabeled puromycin established that resistance was due to a lowered uptake of puromycin and that an inverse relationship existed between resistance level and uptake rate.
AB - Resistance to 10 μg/ml of puromycin has been analyzed in V79 Chinese hamster cells. Clones that were isolated in 10 μg/ml of puromycin and subsequently cultivated in its absence consistently lost their resistance. One clone was analyzed in detail by recloning in the presence and absence of puromycin, and it was found that non-puromycin cultivated sublones also lost their resistance and regained inhibition profiles similar to the V79 parent. Reconstruction experiments between sensitive and resistant cells demonstrated that the yield of mutants was not affected by metabolic cooperation. The mutation rate was calculated to be 1 × 10-7 per cell per generation, and was the same within the limits of statistical error in a colchicine-produced polyploid derivative of the V79 line. Although a number of resistant clones were found to have polyploid karyotypes, the polyploid V79 line was not more resistant to puromycin, nor did it possess a higher frequency of puromycin resistant cells. Studies employing radiolabeled puromycin established that resistance was due to a lowered uptake of puromycin and that an inverse relationship existed between resistance level and uptake rate.
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U2 - 10.1016/0027-5107(80)90098-6
DO - 10.1016/0027-5107(80)90098-6
M3 - Article
C2 - 7360149
AN - SCOPUS:0018835308
SN - 0027-5107
VL - 69
SP - 333
EP - 346
JO - Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
JF - Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
IS - 2
ER -