Part:BBa_K1979004

GFP-PBP5

This device codes for the GFP-PBP5 fusion protein.

Figure1. PCR check for BBa_K1979004. GFP sequence is magnified by primers, with a size of 176 bps (red box).

Figure2. M: Marker; 1: the initial E.coli suspension; 2: deposit of E.coli after centrifugation; 3: deposit of disrupted E.coli; 4: supernatant of disrupted E.coli; 5: W1; 6: W2; 7: W3; 8-11: Elute.

The PBP-GFP was purified through Ni-chelating affinity chromatography. The target protein, PBP-GFP, is marked in the red box, has a molecular weight of 60kDa. As demonstrated in channel 1-4 that protein is identified at the position of 60kDa, PBP-GFP is expressed in the E.coli; as demonstrated in channel 8-11 that the target protein is found in all 4 elutes, we have successfully purified PBP-GFP from the E.coli.

Figure3. M: Marker 1-13: protein through Q Sepharose Fast Flow.

The target protein was purified through Q Sepharose Fast Flow. Proteins in well 3-5 have clear marks at 60kDa, indicating our construction can express the GFP-PBP we desire.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 135
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1892
  Illegal BamHI site found at 168
  Illegal XhoI site found at 2100
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1206
  Illegal AgeI site found at 1808
  
• 1000
  COMPATIBLE WITH RFC[1000]