الفهرس 
LITERATURE REVIEW
Factors controlling expression in E.coliOnly 14 pages are availabe for public view
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Abstract
Production of therapeutic proteins in prokaryotic
system, Escherichia coli (E.coli); has been recognized as an
effective stage for production of recombinant human
interferon gamma (rhIFN-γ). Modification of rhIFN-γ
expression is a line that can be positively employed for
increasing the yield of production through optimization of
induction conditions in shaker flasks to be applied onto batch
culture of recombinant E. coli. Hence, in this research
induction conditions for the over-production of rhIFN-γ
including type of media, pH, type and amount of inducer were
optimized. The factors considered for optimized conditions of
the recombinant E.coli were the use of growth medium LB,
neutral pH7 and the inducer (lactose) at final concentration
2mM. These factors found to be useful in batch process
development. The cell density was reached to 7gm/L wet cell
weight after 12h of batch fermentation.
Commonly, the recombinant proteins were produced in
E. coli as insoluble aggregates called inclusion bodies (IBs). A
method for purification and refolding of rhIFN-γ from IBs has
been designated. It includes solubilization of IBs in 8M
guanidinium hydrochloride; refolding of rhIFN-γ by rapid
dilution method; and protein purification by Hitrap Q XL
strong anion chromatography. The rhIFN-γ obtained has been
characterized by immunogenicity against the hIFN-γ
antibodies. The specific activity of purified rhIFN-γ was 1.87
x 107 IU/mg compared to standard rhIFN-γ via new MxA
reporter gene assay which depends on identifying MxA
mRNA level using real time PCR. The rhIFN-γ increases
expression of the MxA gene product in direct relation to the
dose of rhIFN-γ in IU.