Construction of a Baculovirus vector containing A subunit of Shiga toxin for protein delivery.

BACKGROUND AND OBJECTIVES

Baculovirus can be used as a vector in gene delivery system. Viral envelope of baculovirus would display expressed protein/peptide and it could render as a potential vaccine delivery system. In this regard, the gene coding for A subunit of shiga toxin (StxA) from Escherichia coli (E. coli) strain was cloned in a baculovirus expression system. StxA subunit has the ability to inhibit protein synthesis and this ability applied in cancer therapy. In this study, expression of StxA in baculovirus as a protein delivery system was assessed in vitro.

MATERIAL AND METHODS

StxA gene was cloned in pTriEx™ multisystem expression vector. This vector enables the protein expression in multisystem, E. coli and baculovirus. This construct was used to express the gene in E. coli and baculovirus. The construct containing StxA gene was made in baculovirus and expression was confirmed, then baculovirus expressing STXA transfect HeLa cells.

RESULTS

The expression of STXA peptide (32kDa) was confirmed by SDS-PAGE and western blotting in both expression systems. The A subunit challenge to human cell Lines was applied as a delivery system by baculoviruses. On the other hand, the inhibition of cell proliferation was also demonstrated by baculovirus containing STXA subunit.

CONCLUSION

STXA peptide expression in baculovirus was shown in E. coli and baculovirus expression system. Furthermore, it was shown that A subunit of Shiga toxin delivered by baculovirus can inhibit cell proliferation in HeLa cells and leading to cell death. Therefore, this prototype system could be a promising model for in vivo cancer therapy and targeted protein delivery system.