Coding

Part:BBa_K4849026:Design

Designed by: Devansh Kumar and Catarina Almeida   Group: iGEM23_Edinburgh   (2023-10-11)


CymA - membrane-associated tetraheme c-type cytochrome


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

We wanted to Express CymA in conjugation with the MTR compelx. Based on the current understanding, the electrons extracted from the oxidation of a substrate are ultimately transferred to the periplasmic decaheme cytochrome MtrA during anaerobic respiration. Subsequent electron transfer to the outer membrane β-barrel protein MtrB places MtrA in direct contact with MtrC. MtrC is re-oxidised by reducing terminal electron acceptors, such as electrodes, via direct or mediated electron transfer. We extracted the Gene from Shewanella oneidensis MR-1 (NCBI:txid211586) genomci DNA using Primers that introduced cynogate assmbly system overhangs (refer IGEM Edinburgh 2023 wiki) CymA Forward Primer: CAGTgaagacATaATGAACTGGCGTGCACTA CymA Reverse Primer: CAGTgaagacATaagcTTATCCTTTTGGATAGGGGTG The overhangs were designed to be cut by BbsI - this would allow introduction into the Cynogate Lv0 acceptor vector Gel Showing CymA extracted by PCR:



Source

Shewanella oneidensis MR-1 (NCBI:txid211586)

References