pcr-mediated expression of the human gm-csf gene in escherichia coli

Authors

مانا علومی

mana oloomi سعید بوذری

saeid bouzari

veladimir o. rechinsky

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(+) ex pression 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.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

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 inducti...

full text

PURIFICATION AND CHARACTERIZATION OF THE CLONED HUMAN GM-CSF GENE EXPRESSED IN ESCHERICHIA COLI

The human granulocyte-macrophage colony stimulation factor (hGM-CSF) gene was cloned in the pET 23a( +) expression vector under the control of strong bacteriophage T7 transcription and translation signals. The hGM-CSF gene was transferred into E. coli strainBL21 (DE3)pLysS andIPTG was used for induction of GM-CSF gene. Production of the target protein was obtained as revealed by ELISA and ...

full text

purification and characterization of the cloned human gm-csf gene expressed in escherichia coli

the human granulocyte-macrophage colony stimulation factor (hgm-csf) gene was cloned in the pet 23a( +) expression vector under the control of strong bacteriophage t7 transcription and translation signals. the hgm-csf gene was transferred into e. coli strainbl21 (de3)plyss andiptg was used for induction of gm-csf gene. production of the target protein was obtained as revealed by elisa and weste...

full text

RT-PCR MEDIATED CLONING OF HUMAN GROWTH HORMONE GENE AND I TS EXPRESSION IN E. coli

Human growth hormone (hGH) genomic sequence containing 5 exons and 4 introns was cloned in pcDNA-3 and the constructed plasmid was subsequently used for transfection ofNlli-3T3 cell line using lipofection technique. Expression of hGH in stably transfected cells was assayed using ELISA. Total RNA was extracted from transfected cells and hGH cDNA was amplified by RT-PCR using specific primers...

full text

rt-pcr mediated cloning of human growth hormone gene and i ts expression in e. coli

human growth hormone (hgh) genomic sequence containing 5 exons and 4 introns was cloned in pcdna-3 and the constructed plasmid was subsequently used for transfection ofnlli-3t3 cell line using lipofection technique. expression of hgh in stably transfected cells was assayed using elisa. total rna was extracted from transfected cells and hgh cdna was amplified by rt-pcr using specific primers. th...

full text

The Expression of Human Granulocyte Macrophage Colony Stimulating Factor by Heat-Induction in Escherichia coli

A self-regulated high-copy number plasmid containing chloramphenicol resistant gene, for the production of recombinant proteins under the regulation of bacteriophage ?pL promoter, was constructed. The designed 5024 base pair expression plasmid contained a heat sensitive repressor cI857 coding gene to regulate the function of ?pL promoter under heat shock induction. Using the constructed vector,...

full text

My Resources

Save resource for easier access later


Journal title:
iranian biomedical journal

جلد ۲، شماره ۱، صفحات ۲۱-۲۵

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023