| dc.description.abstract | Recombinant human Interferon-β-1b (IFN-β-1b) is a non-glycosylated IFN-β that
was found to be effective against multiple sclerosis. More recently, expression of
IFN-β-1b in Escherichia coli (E. coli) has been reported; however, problems due
to low yields and poor stability were described. This study was aim to clone and
express of recombinant IFN-β-1b in E. coli using a novel synthetic gene with
preferred codon usage of E. coli under the control of T7 promoter. Protein
expression was induced with Isopropyl-thiogalactoside (IPTG) and lactose. The
level of expression was analyzed by SDS-PAGE, Western blot, Bradford and
ELISA. The effect of two factors including inducer concentration, and length of
the induction on the expression of IFN-β-1b was investigated. As a result, we
have successfully cloned the E. coli BL21 (DE3) stain that harbors gene encoding
for rhIL-29. Results from SDS-PAGE, Western blot and ELISA show that IPTG of
1mM and induction duration of 6h had more effect on IFN-β-1b production. IFN-
β-1b expression with the above condition yielded 17.39% of the total cell
protein. Besides, under the induction of 0.25% lactose and 6 hours of incubation,
IFN-β-1b production also achieved a high efficiency 13.4% of total cell protein,
which promises for an effective way for industrial manufacture.
Keywords: Expression, Interferon-β-1b, E. coli BL21 (DE3), T7 promoter,
induction. | en_US |
