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Team:USTC/Project/protein/protein
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= ''Part Ⅱ: Fusion Proteins for Transportation into BMC'' = | = ''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 ( | + | 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. | ||
Revision as of 11:54, 25 October 2010
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.
