Part:BBa_K258008

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

[1]PrtDEF: ABC transporter of Erwinia chrysanthemi for PrtA
[2]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]