Transfection with a cDNA encoding a Ser(31) or Ser(34) mutant human dihydrofolate reductase into Chinese hamster ovary and mouse marrow progenitor cells confers methotrexate resistance Journal Article

  

Authors:	Banerjee, D.; Schweitzer, B. I.; Volkenandt, M.; Li, M. X.; Waltham, M.; Mineishi, S.; Zhao, S. C.; Bertino, J. R.
Article Title:	Transfection with a cDNA encoding a Ser(31) or Ser(34) 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. © 1994.
Keywords:	sequence analysis; nonhuman; methotrexate; phenotype; animal; mice; gene amplification; bone marrow; serine; drug resistance; transfection; gene transfer; molecular sequence data; genetic transfection; cell culture; dna amplification; stem cells; base sequence; plasmids; mutagenesis, site-directed; bone marrow cell; tetrahydrofolate dehydrogenase; colony-forming units assay; blotting, northern; arginine; genetic markers; complementary dna; retroviridae; mutant; dna, complementary; pcr; gene transfer techniques; cho cell; cho cells; blotting, southern; cloning vector; human; priority journal; article; support, non-u.s. gov't; support, u.s. gov't, p.h.s.; hamsters
Journal Title:	Gene
Volume:	139
Issue:	2
ISSN:	0378-1119
Publisher:	Elsevier Science, Inc.
Date Published:	1994-02-25
Start Page:	269
End Page:	274
Language:	English
DOI:	10.1016/0378-1119(94)90768-4
PROVIDER:	scopus
PUBMED:	8112617
DOI/URL:	https://www.scopus.com/inward/record.uri?eid=2-s2.0-0028230635&doi=10.1016%2f0378-1119%2894%2990768-4&partnerID=40&md5=114f9ea2498d89b9f5f08e5b406c0a57
Notes:	Export Date: 14 January 2019 -- Article -- Source: Scopus

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