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

Authors

  • MANA OLOOMI From the Molecular Biology Unit, Pasteur Institute of Iran, Tehran I.R. Iran.
  • SAEID BOUZARI
  • VLADIMIR O RECHINSKY
Abstract:

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 Western blot analysis. The produced 'h:GM-CSF was purified by immunoaffinity chromatography. The dot blot positive fractions were assayed for biological activity and it was shown that the expressed GM-CSF is active.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

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

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

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(+) 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 inducti...

full text

Purification and characterization of glutathione reductase encoded by a cloned and over-expressed gene in Escherichia coli.

An expression vector, pKGR, for the gor gene from Escherichia coli encoding glutathione reductase was constructed by subcloning of an AvaII fragment of the Clarke & Carbon bank plasmid pGR [Greer & Perham (1986) Biochemistry 25, 2736-2742] into the plasmid pKK223-3. The expression of glutathione reductase from the plasmid pKGR was found to have been successfully placed under the control of the ...

full text

Purification and characterization of human H-ras proteins expressed in Escherichia coli.

The full-length normal and T24 mutant human H-ras proteins and two truncated derivatives of the T24 mutant were expressed efficiently in Escherichia coli. The proteins accumulated to 1 to 5% of total cellular protein, and each was specifically recognized by anti-ras monoclonal antibodies. The two full-length proteins as well as a carboxyl-terminal truncated derivative (deleted for 23 amino acid...

full text

Purification and biochemical characterization of a soluble human interferon gamma receptor expressed in Escherichia coli.

We purified and characterized a soluble human interferon gamma receptor expressed in Escherichia coli. The soluble receptor comprises the amino acids 15-246 of the encoded protein (Aguet, M., Dembic, Z., and Merlin, G. (1988) Cell 55, 273-280) and was purified from large scale fermentations through four chromatographic steps with an overall recovery of 28%. The refolded soluble receptor shows s...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 12  issue 4

pages  353- 357

publication date 1999-02

By following a journal you will be notified via email when a new issue of this journal is published.

Keywords

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023