Part:BBa_K519003

Inducible BMC shell protein generator with GFP fused pduP1~18 tag

P(R0011)-RBS(B0034)-pduABJKNU-Term(B0015)-P(J23102)-RBS(B0034)-pduP1~18-GFP-Term(B0015)

BBa_K519002 was combined to BBa_K317030 a PduP protein with the N-terminal ends known to be needed to be taken into Pdu micro bacterial compartment(BMC), fused to green fluorescent protein we construted in iGEM2010.

The pdu operon from Citrobacter freundii encodes six shell proteins (PduA, -B, -J, -K, -N, and -U) of the propanediol utilization microcompartment.

PduP (propionaldehyde dehydrogenase) protein with the N-terminal ends is known to be needed to be taken into Pdu BMC. The short N-terminal peptide is necessary for packing enzymes into the pdu BMC. It has been reported that the deletion of 10 or 14 amino acids from the N terminus of the enzyme resulted in impaired packaging. On the other hand, fusion of the 18 N-terminal amino acids from PduP to GFP, GST, or maltose-binding protein resuted them to be encapsulated in BMC. The PduP1~18 protein can be fused to an aiming compound which can then be brought into the BMC.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 2821
  Illegal NheI site found at 2844
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1755
  Illegal BamHI site found at 3449
  Illegal XhoI site found at 3350
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 2539
  
• 1000
  COMPATIBLE WITH RFC[1000]