ESCHERICHIA COLI STRAIN BL21: CLONING AND EXPRESSION OF AN OPTIMIZED INTERFERON ALPHA 2B (DE3)

D. S.  Alrseetmiwe; A. A.  Almayah; A. A.  Nasser; M.H. Alnussairi; H.  Alizadeh; F. A .  Mehrzi

doi:10.59807/jlsar.v1i2.15

Authors

D. S. Alrseetmiwe Department of Field Crop, College of Agriculture, University of Basrah, Iraq.
https://orcid.org/0009-0001-3430-856X
A. A. Almayah College of Pharmacy, University of Basrah, Iraq.
https://orcid.org/0000-0002-4254-4319
A. A. Nasser Department of Field Crop, College of Agriculture, University of Basrah, Iraq.
https://orcid.org/0000-0002-1184-6728
M.H. Alnussairi College of Pharmacy, University of Maysan, Iraq.
H. Alizadeh Department of Field Crop, College Agriculture, Tehran University, Iran.
https://orcid.org/0000-0001-8138-0786
F. A . Mehrzi Department of Field Crop, College Agriculture, Tehran University, Iran.

https://doi.org/10.59807/jlsar.v1i2.15

Keywords:

Recombinant Protein, Bacterial Expression System, IPTG Induction, Western Blotting

Abstract

Interferon alpha 2b gene (INF α2b) as a protein with antiviral and antitumor activities is potentially a valuable therapeutic protein to work on. Prior to having a large-scale production of the target protein, it is recommended to examine it on an experimental scale, so that a bacterial host could be a proper choice as it leads us to a deep insight into the subject. In this research, the INF α2b sequence was obtained from the NCBI gene data bank, and after optimization, it was subjected to be cloned and expressed in pET28a+. In order to primary examination of the target protein, Escherichia coli was considered a prokaryotic expression system. IPTG induction of the protein in bacteria cells containing the construct pET: IFN, followed by resolving total proteins through SDS-PAGE. The expected size of the investigated protein, about 24kDa, was observed through gel separation. Further assessment via western blotting confirmed the successful expression of IFN α2b.

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