Abstract
Oral vaccine is one of the most potential strategies for gastrointestinal infection due to significant advantages over parenteral vaccines. In order to overcome exsiting hurdles in oral vaccine, namely oral tolerance and antigen dispersion, gut-associated M cell is a target for vaccine delivery. Cellular prion protein (PrPC), an M cell receptor, was experimentally proven to interact with heat shock protein Hsp60. In the previous in sillico studies, two Hsp60-deprived peptides have been predicted to be key-players in this interaction. Therefore, providing PrPC for in vitro binding assay with Hsp60 is desired to confirm the binding activity of the two predicted peptides. In this study, murine PrPC-encoding gene (mPrPc) was cloned into pET-gst to generate recombinant vector namedly pET-gst-mPrPc. Next, the vector was transformed into expression host Escherichia coli (E. coli) BL21 (DE3) for protein expression. SDS-PAGE and Western blot probed with anti-GST antibodies showed the protein expressed in inclusion bodies and hence was subsequently solubilized and refolded. After refolding, GST-mPrPC protein was harvested in soluble form with the refolding efficiency reached 88.33%.
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