Proceedings of the National Academy of Sciences · 1985 · 1.1K citations · 39 references
Life SciencesGeneticsMolecular GeneticsGene ExpressionGenetic EngineeringGenomicsAnemiaProtein SynthesisChinese Hamster Ovary Cell ExpressionChinese Hamster Ovary CellsGenomic Library Construction
The human erythropoietin gene was isolated from a genomic phage library using mixed 20‑mer and 17‑mer oligonucleotide probes. The 5.4‑kb HindIII‑BamHI fragment encodes a 27‑aa signal peptide and a 166‑aa mature protein (Mr 18,399) with four 1562‑bp introns and five exons, and its expression in Chinese hamster ovary cells yields biologically active erythropoietin in vitro and in vivo.
The human erythropoietin gene has been isolated from a genomic phage library by using mixed 20-mer and 17-mer oligonucleotide probes. The entire coding region of the gene is contained in a 5.4-kilobase HindIII-BamHI fragment. The gene contains four intervening sequences (1562 base pairs) and five exons (582 base pairs). It encodes a 27-amino acid signal peptide and a 166-amino acid mature protein with a calculated Mr of 18,399. The erythropoietin gene, when introduced into Chinese hamster ovary cells, produces erythropoietin that is biologically active in vitro and in vivo.
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