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

Debabrata Banerjee; Barry I. Schweitzer; Matthias Volkenandt; Ming Xia Li; Mark Waltham; Shin Mineishi; Shi Cheng Zhao; Joseph R. Bertino

doi:10.1016/0378-1119(94)90768-4

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

Debabrata Banerjee, Barry I. Schweitzer, Matthias Volkenandt, Ming Xia Li, Mark Waltham, Shin Mineishi, Shi Cheng Zhao, Joseph R. Bertino

Research output: Contribution to journal › Article › peer-review

32 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	269-274
Number of pages	6
Journal	Gene
Volume	139
Issue number	2
DOIs	https://doi.org/10.1016/0378-1119(94)90768-4
State	Published - Feb 25 1994

All Science Journal Classification (ASJC) codes

Genetics

Access to Document

10.1016/0378-1119(94)90768-4

Cite this

Banerjee, D., Schweitzer, B. I., Volkenandt, M., Li, M. X., Waltham, M., Mineishi, S., Zhao, S. C., & Bertino, J. R. (1994). 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. Gene, 139(2), 269-274. https://doi.org/10.1016/0378-1119(94)90768-4

@article{b8a8a2e69baf4b04aa959274262a6b64,

title = "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",

abstract = "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.",

author = "Debabrata Banerjee and Schweitzer, {Barry I.} and Matthias Volkenandt and Li, {Ming Xia} and Mark Waltham and Shin Mineishi and Zhao, {Shi Cheng} and Bertino, {Joseph R.}",

note = "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.",

year = "1994",

month = feb,

day = "25",

doi = "10.1016/0378-1119(94)90768-4",

language = "English (US)",

volume = "139",

pages = "269--274",

journal = "Gene",

issn = "0378-1119",

publisher = "Elsevier B.V.",

number = "2",

}

Banerjee, D, Schweitzer, BI, Volkenandt, M, Li, MX, Waltham, M, Mineishi, S, Zhao, SC & Bertino, JR 1994, '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', Gene, vol. 139, no. 2, pp. 269-274. https://doi.org/10.1016/0378-1119(94)90768-4

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. / Banerjee, Debabrata; Schweitzer, Barry I.; Volkenandt, Matthias et al.
In: Gene, Vol. 139, No. 2, 25.02.1994, p. 269-274.

Research output: Contribution to journal › Article › peer-review

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.

UR - http://www.scopus.com/inward/record.url?scp=0028230635&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028230635&partnerID=8YFLogxK

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 -

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

Abstract

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this