Protein_Domain

Part:BBa_K371017:Design

Designed by: Yang Zhang   Group: iGEM10_USTC   (2010-10-09)

pduV1-98


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The present BioBrick assembly standard RFC does not support Protein fusion. In our project, we propose a new BioBrick assembly standard BBF RFC53---MetaPart Assembly Standard, which extending RFC 10 to Enable Scarless Protein Fusion with Type IIS Restriction Enzyme EarI and SapI.

In the previous literature，most of research work on pdu BMC(or pdu MCP) focus on Citrobacter Freundii and Salmonella enterica. Taking all of the factors together, we choose Citrobacter Freundii as the orgin of gene in our project. Because the whole genome of Citrobacter Freundii has not been decoded yet, we use the reference sequence sent to NCBI by Martin J.Warren group(GenBank: AM498294.1).

In order to make the new parts compatible to new standard, we firstly check the sequence of pduV, which we get from GenBank(GenBank: AM498294.1), with the new four restriction enzymes site EarI, SapI,SacI and BglII. Luckily, none of these site were find in the 298bp sequence of pduV. Then we use the following primers to get the pduV98 from Citrobacter Freundii genome. The final part consist of cds of pduV98 with meta-prefix and meta-suffix in its sides

forward primer:5'-GTTTCTCTTCAATGAAACGCATAATGCTAATTGGCCCCAG-3'

reverse primer:5'-GTTTCTCTTCAACCTTTGGTAACGAGGCCAATAACGGGTCGG-3'

Source

Citrobactor freudii, the strain was bought from NITE Biological Resource Center (NBRC). NBRC NO. is 12681. History:IFO 12681 <- Ajinomoto Co., Inc. (AJ 2619) <- ATCC 8090 <- C.H. Werkman

References