PCR-mediated Expression of the Human GM-CSF Gene in Escherichia coli

Four exons of the human genomic GM-CSF gene were assembled together using gene splicing by overlap extension (SOE) method. The resulting nucleotide sequence was cloned in the pET23a(+) ex‌pression vector under the control of strong bacteriophage T7 transcription and translation signals. The construct obtained was Transferred into the E. coli strain, BL21(DE3) pLysS and IPTG was used for induction of GM-CSF gene and production of the target protein. one mg of protein per liter of cell culture, was obtained as revealed by ELISA.