Erythropoietin, a glycoprotein synthesized by the kidney , is the primary regulator of erythropoiesis and can stimulate erythroblast proliferation and differentiation. The cloning of erythropoietin has led to the way for production of the recombinant hormone and has been useful for the treatment of anemic patients with kidney failure. Less is known about the erythropoietin receptor which has been recently cloned from the mRNA of murine erythroleukemia cells. We I have isolated a genomic clone containing the human erythropoietin receptor from a human placental library using a 400 bp probe from the 5'coding region of the murine erythropoietin receptor cDNA. The erythropoietin receptor gene is contained within 14 kb of genomic DNA inserted into a lambda phage vector. The coding region is localized within about 6 kb and is interrupted by several intervening sequences ranging in size from 79 bp to I kb. The intervening sequences are rich in Alu-related sequences. Partial sequence analysis of the coding region has enabled us to synthesize several oligonucleotide primers which were useful in generating PCR fragments from cDNA prepared from OMC1 cells, a human erythroleukemia cell line with high levels of the erythropoietin receptor on its surface. With the cloning of the genomic human erythropoietin receptor, it will now possible to study the transcriptional control of the erythropoietin receptor, its specific binding to erythropoietin and the process of signal transduction upon binding of erythropoietin at the cell surface.

Project Start
Project End
Budget Start
Budget End
Support Year
2
Fiscal Year
1990
Total Cost
Indirect Cost
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State
Country
United States
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