Volume 5, Issue 19 (7-2015)
NCMBJ 2015, 5(19): 19-23 |
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Khatami H R, fallah zadeh R, fallah mehrabadi J, Ghafari M D. Expression assessment of recombinant human granulocyte colony stimulating factor (hG-CSF) in E. coli (Origami / BL21). NCMBJ 2015; 5 (19) :19-23
URL: http://ncmbjpiau.ir/article-1-657-en.html
URL: http://ncmbjpiau.ir/article-1-657-en.html
Department of Bioscience and Biotechnology, Malek Ashtar University, Tehran, Iran. , Raminfallah2009@yahoo.com
Abstract: (9997 Views)
Aims and Background: Human granulocyte colony stimulating factor (hG-CSF) is a hematopoietic cytokine that stimulates activation of mature blood cells. Nowadays, recombinant hG-CSF is successfully used for the treatment of neutropenia in patients suffering from cancer of myeloid suppressor, bone marrow transplant, acute myeloid leukemia and acute/severe neutropenia. The aim of this study was the increased expression of hG-CSF and production of the soluble protein with disulfide bonds by E.coli strains (Origami and BL21).
Materials and Methods: The synthesized hG-CSF gene (in pGEM vector) was subcloned into pET23a expression vector and then the recombinant plasmid was transformed into Origami and BL21 hosts. Afterwards, expression levels were compared in two hosts.
Results: Enzymatic digestion results showed the accuracy of the cloning procedures in vector pET23a. Also under the same conditions and after induction of different hosts with IPTG, best hG-CSF protein expression was observed in strain Origami E.coli.
Conclusion: The results showed that E.coli (Origami) is appropriate strain for the expression of human G-CSF and this strain can be used as suitable host for the expression of this protein in higher scale.
Materials and Methods: The synthesized hG-CSF gene (in pGEM vector) was subcloned into pET23a expression vector and then the recombinant plasmid was transformed into Origami and BL21 hosts. Afterwards, expression levels were compared in two hosts.
Results: Enzymatic digestion results showed the accuracy of the cloning procedures in vector pET23a. Also under the same conditions and after induction of different hosts with IPTG, best hG-CSF protein expression was observed in strain Origami E.coli.
Conclusion: The results showed that E.coli (Origami) is appropriate strain for the expression of human G-CSF and this strain can be used as suitable host for the expression of this protein in higher scale.
Keywords: granulocyte colony stimulating factor (hG-CSF), Recombinant Protein, optimization of codon, E.coli Expression strains
Type of Study: Research Article |
Subject:
Cellular and molecular
Received: 2015/07/16 | Accepted: 2015/07/16 | Published: 2015/07/16
Received: 2015/07/16 | Accepted: 2015/07/16 | Published: 2015/07/16
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