中国生物工程杂志

To achieve high level expression of human transferring (hTF) and investigate the possible mechanism of the hTF expression, rabbit transferrin promoter (RP promoter) was cloned, and a hTF recombinant expression vector (δpcDNA3.1(-)-RP-hTF), was constructed in which hTF DNA was driven by RP promoter. The transient transfection experiments achieved a high hTF expression in BRL-3A and HC-11 cell lines, and two stably integrated cell clones were also screened out. Various concentrations of 5-aza-dC were added to the cell culture medium for the investigation of the effect on hTF expression. Real-time RT-PCR analysis showed that the RP promoter directed an efficient hTF expression, and 5-aza-dC increased hTF expression in a proportional manner. However, the extent of the increase in hTF expression differed between individual clones, suggesting that TF expression may relate to gene methylation.