Difference between revisions of "Part:BBa K1979004"
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<p class="text">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.<br/><br/>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.</p>
 
<p class="text">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.<br/><br/>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.</p>
 
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<img src="https://static.igem.org/mediawiki/parts/9/92/T--SDSZ_China--GFP-PBP_part_3.jpg" style= "width:60%;"/">
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<p class="text">Figure3. M: Marker 1-13: protein through Q Sepharose Fast Flow.<br/><br/>The target protein was purified through Q Sepharose Fast Flow. Proteins in channel 3-5 have clear marks at 60kDa, indicating our construction can express the GFP-PBP we desire.</p>
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<p class="text">Figure3. M: Marker 1-13: protein through Q Sepharose Fast Flow.<br/><br/>The target protein from channel 8-11 in figure 2 was purified through Q Sepharose Fast Flow. Proteins in channel 3-5 have clear marks at 60kDa, indicating our construction can express the GFP-PBP we desire.</p>
 
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Revision as of 17:00, 19 October 2016


GFP-PBP5

This device codes for the GFP-PBP5 fusion protein.

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]

Figure1. PCR check for BBa_K1979004. GFP sequence is magnified by primers, with a size of 716 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 from channel 8-11 in figure 2 was purified through Q Sepharose Fast Flow. Proteins in channel 3-5 have clear marks at 60kDa, indicating our construction can express the GFP-PBP we desire.