Part Ⅱ: Fusion Proteins for Transportation into BMC

It is reported that a short N-terminal peptide is necessary and sufficient for packing enzymes into the lumen of the BMC. Fusion of the 14, 18 or 64 N-terminal amino acids from PduP to GFP or RFP resulted in their encapsulation within BMCs. Each fusion protein (PduP[1-14]- GFP, PduP[1-18]- GFP, PduP[1-64]- GFP) was produced by the new standard. Our new standard, as it is stated above, consists of three restriction enzyme cut sites --- SacⅠ(CCTCG)，EarⅠ(CTCTTC) and SapⅠ(GCTCTTC). The details of enzyme digestion and ligation can be seen in Fig 2a.

When 14/18/64 amino acids from the N terminus of PduP were fused to GFP, the fusion protein was readily detected by Western blotting. These results, in conjunction with the above studies, indicate that a short region of the N terminus of PduP is necessary and sufficient for packing proteins to the lumen of the BMC.