A new vector using the human multidrug resistance gene as a selectable marker enables overexpression of foreign genes in eukaryotic cells.
A new vector, pSK1.MDR, has been constructed for expressing nonselectable genes in eukaryotic cells. The vector uses the human multidrug resistance gene, MDR1, as a dominant selectable marker and contains an additional transcription unit plus a unique SalI cloning site for inserting nucleotide sequences to be expressed. To test this expression system, a cDNA (IL2R) for the 55-kDa interleukin-2 receptor was inserted into the SalI site, and the resulting plasmid was transfected into NIH3T3 cells. Cells which acquired the MDR1 gene were selected with colchicine, and cells with high levels of MDR1 expression were selected by growth in increasing concentrations of the drug. Drug resistant cells also expressed the cotransferred, nonselected IL2R gene, and its expression was increased to 740,000 receptors per cell by growing cells in high concentrations of colchicine. The MDR1 system represents a very efficient method for synthesizing large amounts of protein in a wide variety of eukaryotic cells.
Kane
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National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Reinhard
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DH
Fordis
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Pastan
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I
Gottesman
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D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000818:Animals; D002460:Cell Line; D003078:Colchicine / Q000494:pharmacology; D004351:Drug Resistance / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D005057:Eukaryotic Cells / Q000378:metabolism; D005434:Flow Cytometry; D005784:Gene Amplification; D015870:Gene Expression; D005819:Genetic Markers; D005822:Genetic Vectors; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics*; D010957:Plasmids; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D015375:Receptors, Interleukin-2 / Q000235:genetics; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D005819:Genetic Markers; D008562:Membrane Glycoproteins; D015375:Receptors, Interleukin-2; D003078:Colchicine