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פותח על ידי קלירמאש פתרונות בע"מ -
Isolation of methotrexate-resistant cell lines in Petunia hybrida upon stepwise selection procedure
Year:
1984
Source of publication :
Plant Molecular Biology
Authors :
ברג, רבקה
;
.
פלג, נעמי
;
.
פרל, מאיר
;
.
Volume :
3
Co-Authors:
Barg, R., Department of Plant Genetics and Breeding, Institute of Field and Garden Crops, Agricultural Research Organization, Bet-Dagan, 50-250, Israel
Peleg, N., Department of Plant Genetics and Breeding, Institute of Field and Garden Crops, Agricultural Research Organization, Bet-Dagan, 50-250, Israel
Perl, M., Department of Plant Genetics and Breeding, Institute of Field and Garden Crops, Agricultural Research Organization, Bet-Dagan, 50-250, Israel
Beckmann, J.S., Department of Plant Genetics and Breeding, Institute of Field and Garden Crops, Agricultural Research Organization, Bet-Dagan, 50-250, Israel
Facilitators :
From page:
303
To page:
311
(
Total pages:
9
)
Abstract:
Cell suspensions of Petunia hybrida were subjected to a selection procedure in which the concentration of the selective agent, methotrexate (MTX), was gradually elevated. In mammalian cells, this procedure frequently results in MTX-resistant mutants due to amplification of the gene coding for dihydrofolate reductase (DHFR), the target protein of MTX. Five suspension lines were isolated, with degrees of resistance ranging from 10 to 500 μM MTX (in wild type the LD99.9 is 0.2 μM). MTXR phenotypes were unstable, as manifested by the loss of resistance upon prolonged growth in the absence of drug. All of the mutants also exhibited high values of MTX-binding protein (60- to 400-fold higher than that of the wild type), which declined to intermediate values upon MTX withdrawal. Finally, cellular extracts from all of the mutants also showed high specific staining of DHFR-activity in gels. The results suggest that the resistance of MTX in these plant cell-lines is mediated by the elevation of the amounts of DHFR, probably as a consequence of gene amplification. © 1984 Martinus Nijhoff/Dr W. Junk Publishers.
Note:
Related Files :
Dihydrofolate reductase
gene amplification
MIX-resistance
petunia cell cultures
stepwise selection
עוד תגיות
תוכן קשור
More details
DOI :
10.1007/BF00017784
Article number:
Affiliations:
Database:
סקופוס
Publication Type:
מאמר
;
.
Language:
אנגלית
Editors' remarks:
ID:
20369
Last updated date:
02/03/2022 17:27
Creation date:
16/04/2018 23:35
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Scientific Publication
Isolation of methotrexate-resistant cell lines in Petunia hybrida upon stepwise selection procedure
3
Barg, R., Department of Plant Genetics and Breeding, Institute of Field and Garden Crops, Agricultural Research Organization, Bet-Dagan, 50-250, Israel
Peleg, N., Department of Plant Genetics and Breeding, Institute of Field and Garden Crops, Agricultural Research Organization, Bet-Dagan, 50-250, Israel
Perl, M., Department of Plant Genetics and Breeding, Institute of Field and Garden Crops, Agricultural Research Organization, Bet-Dagan, 50-250, Israel
Beckmann, J.S., Department of Plant Genetics and Breeding, Institute of Field and Garden Crops, Agricultural Research Organization, Bet-Dagan, 50-250, Israel
Isolation of methotrexate-resistant cell lines in Petunia hybrida upon stepwise selection procedure
Cell suspensions of Petunia hybrida were subjected to a selection procedure in which the concentration of the selective agent, methotrexate (MTX), was gradually elevated. In mammalian cells, this procedure frequently results in MTX-resistant mutants due to amplification of the gene coding for dihydrofolate reductase (DHFR), the target protein of MTX. Five suspension lines were isolated, with degrees of resistance ranging from 10 to 500 μM MTX (in wild type the LD99.9 is 0.2 μM). MTXR phenotypes were unstable, as manifested by the loss of resistance upon prolonged growth in the absence of drug. All of the mutants also exhibited high values of MTX-binding protein (60- to 400-fold higher than that of the wild type), which declined to intermediate values upon MTX withdrawal. Finally, cellular extracts from all of the mutants also showed high specific staining of DHFR-activity in gels. The results suggest that the resistance of MTX in these plant cell-lines is mediated by the elevation of the amounts of DHFR, probably as a consequence of gene amplification. © 1984 Martinus Nijhoff/Dr W. Junk Publishers.
Scientific Publication
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