[Synthesis and cloning of the whole human erythropoietin (EPO) gene].
A 600 bp synthetic erythropoietin (EPO) gene encoding all 166 amino acids of the EPO protein and 27 amino acids of the signal peptide has been constructed. The whole gene was divided into three large fragments consisting of a total of 32 oligonucleotides. These oligonucleotides were synthesized by the solid-phase phosphor-amidite method and ligated into three large fragments. These latter three were separately cloned into vector M13mp19 and then transformed into E. coli JM 103. Positive clones were screened with 32P-labeled probes. The sequences of the fragments were confirmed by DNA sequencing, and the sequence of the whole synthetic EPO gene was confirmed by enzymatic digestion and sequencing. The results indicated that the nucleotide sequence of the synthetic EPO gene is identical to that of the original.
Cai
Y
Y
Institute of Haematology, Tianjin.
D003001:Cloning, Molecular; D004247:DNA / Q000138:chemical synthesis / Q000235:genetics; D004921:Erythropoietin / Q000138:chemical synthesis / Q000235:genetics*; D009841:Oligonucleotides / Q000138:chemical synthesis; D010446:Peptide Fragments / Q000138:chemical synthesis
D009841:Oligonucleotides; D010446:Peptide Fragments; D004921:Erythropoietin; D004247:DNA