TY - JOUR
T1 - Transfection with a cDNA encoding a Ser31 or Ser35 mutant human dihydrofolate reductase into Chinese hamster ovary and mouse marrow progenitor cells confers methotrexate resistance
AU - Banerjee, Debabrata
AU - Schweitzer, Barry I.
AU - Volkenandt, Matthias
AU - Li, Ming Xia
AU - Waltham, Mark
AU - Mineishi, Shin
AU - Zhao, Shi Cheng
AU - Bertino, Joseph R.
N1 - Funding Information:
This work was funded by NIH grant CA08010, a Bristol Myers research Grant and by a Cancer Center Support Grant NCI-P30-CA-08748. We thank Laurie Weinstein for excellent technical assistance. M.V. was supported by grant Vo 415/1-l of the Deutsche Forschungsmeinschaft. J.R.B. is an American Cancer Society Professor of Medicine and Pharmacology.
PY - 1994/2/25
Y1 - 1994/2/25
N2 - Chinese hamster ovary (CHO) DHFR- cells were converted into the DHFR+ phenotype when they were transfected with a mammalian expression vector carrying human dihydrofolate reductase-encoding cDNAs (DHFR) containing a Ser31 or a Ser34 mutation. Furthermore, transfection of these mutants into wild-type CHO cells resulted in resistance to high levels of methotrexate (MTX), indicating that these human variants can act as dominant selectable markers. Southern blot analysis and polymerase chain reaction amplifications confirmed that the transfected plasmids were integrated into the CHO DNA. Gene copy number analysis revealed that both the Ser31 and the Ser34 mutants are amplifiable when grown in increasing concentrations of MTX. Retrovirus-mediated gene transfer of the Ser31 mutant into mouse marrow progenitor cells also resulted in MTX-resistant CFU-GM (colony-forming unit-granulocyte macrophage) cells.
AB - Chinese hamster ovary (CHO) DHFR- cells were converted into the DHFR+ phenotype when they were transfected with a mammalian expression vector carrying human dihydrofolate reductase-encoding cDNAs (DHFR) containing a Ser31 or a Ser34 mutation. Furthermore, transfection of these mutants into wild-type CHO cells resulted in resistance to high levels of methotrexate (MTX), indicating that these human variants can act as dominant selectable markers. Southern blot analysis and polymerase chain reaction amplifications confirmed that the transfected plasmids were integrated into the CHO DNA. Gene copy number analysis revealed that both the Ser31 and the Ser34 mutants are amplifiable when grown in increasing concentrations of MTX. Retrovirus-mediated gene transfer of the Ser31 mutant into mouse marrow progenitor cells also resulted in MTX-resistant CFU-GM (colony-forming unit-granulocyte macrophage) cells.
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U2 - 10.1016/0378-1119(94)90768-4
DO - 10.1016/0378-1119(94)90768-4
M3 - Article
C2 - 8112617
AN - SCOPUS:0028230635
SN - 0378-1119
VL - 139
SP - 269
EP - 274
JO - Gene
JF - Gene
IS - 2
ER -