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

被引：34

作者：

BANERJEE, D ^{[1
]}

SCHWEITZER, BI ^{[1
]}

VOLKENANDT, M ^{[1
]}

LI, MX ^{[1
]}

WALTHAM, M ^{[1
]}

MINEISHI, S ^{[1
]}

ZHAO, SC ^{[1
]}

BERTINO, JR ^{[1
]}

机构：

[1] MEM SLOAN KETTERING CANC CTR,MOLEC PHARMACOL & EXPTL THERAPEUT PROGRAM,NEW YORK,NY 10021

来源：

GENE | 1994年 / 139卷 / 02期

关键词：

GENE TRANSFER; DNA AMPLIFICATION; PCR; SEQUENCE ANALYSIS;

D O I：

10.1016/0378-1119(94)90768-4

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

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 Ser(31) or a Ser(34) 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 Ser(31) and the Ser(34) mutants are amplifiable when grown in increasing concentrations of MTX. Retrovirus-mediated gene transfer of the Ser(31) mutant into mouse marrow progenitor cells also resulted in MTX-resistant CFU-GM (colony-forming unit-granulocyte macrophage) cells.

引用

页码：269 / 274

页数：6

共 22 条

[1]

Blakley R. L., 1984, FOLATES PTERINES, P191

[2] SINGLE-STEP METHOD OF RNA ISOLATION BY ACID GUANIDINIUM THIOCYANATE PHENOL CHLOROFORM EXTRACTION [J].

CHOMCZYNSKI, P ;

SACCHI, N .

ANALYTICAL BIOCHEMISTRY, 1987, 162 (01) :156-159

[3]

COREY CA, 1990, BLOOD, V75, P337

[4]

HABER DA, 1981, J BIOL CHEM, V256, P9501

[5] CONSTRUCTION OF A DOMINANT SELECTABLE MARKER USING A NOVEL DIHYDROFOLATE-REDUCTASE [J].

HUSSAIN, A ;

LEWIS, D ;

YU, MH ;

MELERA, PW .

GENE, 1992, 112 (02) :179-188

[6] SYSTEMIC RESISTANCE TO METHOTREXATE IN TRANSGENIC MICE CARRYING A MUTANT DIHYDROFOLATE-REDUCTASE GENE [J].

ISOLA, LM ;

GORDON, JW .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (24) :9621-9625

[7] UPTAKE AND EFFICACY OF TRIMETREXATE (TMQ, 2,4-DIAMINO-5-METHYL-6-[(3,4,5-TRIMETHOXYANILINO)METHYL] QUINAZOLINE), A NON-CLASSICAL ANTIFOLATE IN METHOTREXATE-RESISTANT LEUKEMIA-CELLS INVITRO [J].

KAMEN, BA ;

EIBL, B ;

CASHMORE, A ;

BERTINO, J .

BIOCHEMICAL PHARMACOLOGY, 1984, 33 (10) :1697-1699

[8] COMPARISON OF THE EXPRESSION OF A MUTANT DIHYDROFOLATE-REDUCTASE UNDER CONTROL OF DIFFERENT INTERNAL PROMOTERS IN RETROVIRAL VECTORS [J].

LI, M ;

HANTZOPOULOS, PA ;

BANERJEE, D ;

ZHAO, SC ;

SCHWEITZER, BI ;

GILBOA, E ;

BERTINO, JR .

HUMAN GENE THERAPY, 1992, 3 (04) :381-390

[9]

MCIVOR RS, 1990, NUCLEIC ACIDS RES, V18, P7025, DOI 10.1093/nar/18.23.7025

[10] CRYSTAL-STRUCTURE OF HUMAN DIHYDROFOLATE-REDUCTASE COMPLEXED WITH FOLATE [J].

OEFNER, C ;

DARCY, A ;

WINKLER, FK .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1988, 174 (02) :377-385

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