PCR-mediated expression of the human GM-CSF gene in escherichia coli

ABSTRACT

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