Bacterial surface display of VEGF with the aim of the design of a whole-cell biosensor

Cell surface display technology has recently gained much attention as a powerful tool for biotechnology and biomedical applications. Here, we used the bacterial surface display system for VEGF expression on the surface of E. coli fusing it with the InaK anchoring motif from an ice nucleation protein. Vascular endothelial growth factor (VEGF) is a key regulator of both physiological and pathological angiogenesis. To create the desired construct, after digestion of the amplified VEGF gene and pET۲۱a vector containing the encoding gene of truncated INP (InaK) using the identical restriction enzymes (HindIII/XhoI), the VEGF gene was ligated into the vector. The construct was transformed into E. coli, BL۲۱ (DE۳), followed by the screening of positive colonies and induction of the recombinant bacterial cells with IPTG at ۲۷ °C for ۶ h. After harvesting and lysis of the cells, the bacterial cell wall was analyzed by SDS-PAGE to evaluate VEGF display on the cell surface. The results confirmed the over-expression of VEGF on the surface of bacteria. Such a cell surface display system could be considered as a good platform for the design of whole-cell biosensors.

Keywords:

Cell Surface Display , VEGF , Whole-cell Biosensor