Plasmid
PrtDEF

Part:BBa_K258007

Designed by: Cihan Tastan donated by Jung Hoon Ahn from Korea Advanced Institute of Science and Technology   Group: iGEM09_METU-Gene   (2009-10-10)

Export of recombinant proteins in Escherichia coli using ABC transporter of Erwinia chrysanthemi


EGF-TliA, EGF-LARD1 fusion proteins were secreted when ABC transporter was present. E. coli harboring the PrtDEF of E. chrysanthemi secreted more fusion proteins than without ABC transporter. In fact, in enzyme activation assay(with Hitachi Spectrophotometer) we observed that although our protein -EGF- has disulfide bonds, it was excreted succesfully. In addition, when compearing with lack of ABC domain, we saw that protein excretion into medium (supernatant) is 10-fold more with ABC transporter-PrtDEF-. PrtDEF functioned well at 37°C, the optimum growth temperature for E. coli

Sequence and Features PrtDEF: ABC transporter of Erwinia chrysanthemi for PrtA EGF: Human Epidermal Growth Factor


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 210
    Illegal PstI site found at 793
    Illegal PstI site found at 1140
    Illegal PstI site found at 3055
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 4344
    Illegal PstI site found at 210
    Illegal PstI site found at 793
    Illegal PstI site found at 1140
    Illegal PstI site found at 3055
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1259
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 210
    Illegal PstI site found at 793
    Illegal PstI site found at 1140
    Illegal PstI site found at 3055
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 210
    Illegal PstI site found at 793
    Illegal PstI site found at 1140
    Illegal PstI site found at 3055
    Illegal NgoMIV site found at 4706
    Illegal AgeI site found at 3189
  • 1000
    COMPATIBLE WITH RFC[1000]


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