Cloning of the human erythropoietin exons and their expression in COS-7 cells.
Human erythropoietin exons (hEPO-E) were isolated from a Chinese fetal liver DNA library by using the PCR method with gene recombination. One nucleotide mutation was found in exon 3 leading to the Leu in position 62 being changed to Ser. The hEPO-E coding for the full-length mature protein was inserted into different clone sites of PSV-2-dhfr to create different transferring plasmids (pSV2-dhfr/F1, pSV2/F2, pSV2-dhfr/F3, pSV2-dhfr/F4, pSV2-dhfr/G1, and pSV2-dhfr/G3) which were transfected into COS-7 cells. Results obtained from the comparative experiments indicate that biological activity of hEPO was found in all culture supernatants and its expression level was higher than that of its genome.
Han
F
F
National Laboratory of Molecular Virology and Genetic Engineering, Chinese Academy of Preventive Medicine, Beijing, China.
Wu
S
S
Xue
S
S
Huang
L
L
Ma
X
X
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D003001:Cloning, Molecular; D004247:DNA; D004921:Erythropoietin / Q000235:genetics*; D005091:Exons; D006801:Humans; D008969:Molecular Sequence Data; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D014162:Transfection
D004921:Erythropoietin; D004247:DNA