Part:BBa_K519014
Promoter and RBS with PduP1~18 fused fMT
P(J23117)-RBS(B0034)-PduP1~18-fMT(K190019)
An arsenic binding protein metallothionein fMT was fused to PduP1~1. PduP protein (propionaldehyde dehydrogenase) is an enzyme utilized inside 1,2-propanediol utilization bacterial micro compartment. N-terminal ends of PduP 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.
fMT was added to the BioBrick parts by Team Groningen in iGEM2009, it is a metallothionein (metal binding protein) that can bind to Arsenite(II) and Arsenate(V). It is also known to bind to Cadmium(II). This year for our iGEM project, we evaluated the growth of E. coli in different medium of different Cd(II) concenrations expressing fMT fused to PduP1~18. As a result, we were able to observe that E. coli expressing PduP1~18 fused fMT could tolerate higher concentration of Cd(II) than the wild type E. coli that does not express the metallothionein. We were able to see that fMT still functioned when it was fused with the tag protein.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 153
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 242
- 1000COMPATIBLE WITH RFC[1000]
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