Overexpression of Recombinant Human Beta Interferon (rhINF-β) in Periplasmic Space of Escherichia coli
Authors
Abstract:
Human Interferon β (INF-β) is a member of cytokines family which different studies have shown its immunomodulatory and antiviral activities. In this study an expression vector was designed and constructed for expression of human INF-β-1b either in shake flasks and bench top bioreactor. The designed vector was constructed based upon pET-25b(+) with T7 promoter. Recombinant human beta interferon (rhINF-β) was codon optimized and overexpressed as a soluble, N-terminal pelB fusion protein and secreted into the periplasmic space of Escherichia coli BL21 (DE3). The sugar, Isopropyl-β-D-thiogalactopyranoside (IPTG) was used as a chemical inducer for rhINF-β production in the shake flasks and bench top bioreactor. Timing of beta interferon expression was controlled by using the T7 promoter. The rhINF-β protein was extracted from periplasmic space by osmotic shock treatment and the expression of the beta interferon encoding gene in random selected transformants, was confirmed by western and dot blot methods. The maximum of product formation achieved at the OD600nm = 3.42 was found to be 35 % of the total protein content of the strain which translates to 0.32 g L-1. The constructed vector could efficiently overexpress the rhINF-β into the periplasmic space of E. coli. The obtained yield of the produced rhINF-β was more than previous reports. The system is easily adapted to include other vectors, tags or fusions and therefore has the potential to be broadly applicable to express other recombinant proteins.
similar resources
overexpression of recombinant human beta interferon (rhinf-β) in periplasmic space of escherichia coli
human interferon β (inf-β) is a member of cytokines family which different studies have shown its immunomodulatory and antiviral activities. in this study an expression vector was designed and constructed for expression of human inf-β-1b either in shake flasks and bench top bioreactor. the designed vector was constructed based upon pet-25b(+) with t7 promoter. recombinant human beta interferon ...
full textOverexpression of Recombinant Human Beta Interferon (rhINF-β) in Periplasmic Space of Escherichia coli
Human Interferon β (INF-β) is a member of cytokines family which different studies have shown its immunomodulatory and antiviral activities. In this study an expression vector was designed and constructed for expression of human INF-β-1b either in shake flasks or bench top bioreactor. The designed vector was constructed based upon pET-25b(+) with T7 promoter. Recombinant human beta interferon (...
full textFed-batch Cultivation of Recombinant Escherichia coli Producing Human Interferon-γ Under Controlled Specific Growth Rate
full text
CONSTRUCTION OF RECOMBINANT PLASMIDS FOR PERIPLASMIC EXPRESSION OF HUMAN GROWTH HORMONE IN ESCHERICHIA COLI UNDER T7 AND LAC PROMOTERS
In order to study the periplasmic expression of human growth hormone (hGH) in Escherichia coli, the related cDNA was inserted in two expression plasmids carrying pelB signal peptide, one with lac bacterial promoter and the other with a bacteriophage T7-based promoter. The recombinant plasmids were moved to TG1 and BL21 strains of E. coli, respectively. To induce the expression systems, IPTG and...
full textThe Effect of Heat Shock on Production of Recombinant Human Interferon Alpha 2a (rhIFN α -2a) by Escherichia coli
Recombinant human interferon alpha 2a (rhIFN α -2a) production and cell growth were monitored in a set of genetically modified E. coli strains (MSD1519, MSD1520, MSD 1521, MSD 1522, MSD 1523) producing rhIFN α -2a. The growth was followed at OD 600 nm, changes in cell physiology were detected by pyrolysis mass spectrometry (PyMS) of cell biomass and recombinant protein production was determined...
full textMy Resources
Journal title
volume 13 issue Supplement
pages 151- 160
publication date 2014-02-01
By following a journal you will be notified via email when a new issue of this journal is published.
Hosted on Doprax cloud platform doprax.com
copyright © 2015-2023