ABSTRACT
In the present work, a recombinant Escherichia coli strain was used for the production of interferon α-2b in both shake flask and in bioreactor. The first part of this research was focused on the investigation of the effect of glucose concentration on the kinetics of cell growth, recombinant protein production and acetate formation. In general, glucose supplementation to culture medium enhanced cell growth when added in concentration between 0-20 g/L. Further increase in glucose level reduces biomass production and enhances acetate accumulation in culture. The results clearly demonstrated that maximal interferon production of 27.7 mg/L was achieved in culture supplemented with 20 g/L glucose. Further improvement in recombinant interferon production process was also achieved by scaling up from shake flask to 16-L stirred tank bioreactor. The maximal volumetric interferon production in bioreactor batch culture was 44.5. mg/L after only 6 hours.
ABSTRACT
The study was designed to examine the effect of glucose on the expression of c-myc gene in cultured RINm5F cells. After monolayer culture was established in RPMI 1640 media supplemented with 10% fetal calf serum (FCS), the cells were cultured in various concentrations of glucose and 1 or 10% FCS for another 24 hours. A mRNA was extracted from the cultured cells by a single step method, and Northern analysis was done to detect RNA band. A 0.5 kilobase single band was detected as c-myc mRNA. The expression of c-myc gene mRNA was reduced with increased concentration of glucose with 1% FCS. However, supplementation of 10% FCS abolished the effect of glucose on expression of c-myc gene. These findings suggested that glucose in conjunction with other growth promoting factors played an important role in expression of oncogene and cell growth in RINm5F cells.