Difference between revisions of "Part:BBa K1979004"
Xinyiloowhoo (Talk | contribs) |
|||
| Line 22: | Line 22: | ||
<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
| − | ===Functional Parameters=== | + | ===Functional Parameters===--> |
<partinfo>BBa_K1979004 parameters</partinfo> | <partinfo>BBa_K1979004 parameters</partinfo> | ||
<!-- --> | <!-- --> | ||
Revision as of 00:35, 30 October 2016
GFP-PBP5
This device codes for the GFP-PBP5 fusion protein.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 135 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1892
Illegal BamHI site found at 168
Illegal XhoI site found at 2100 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1206
Illegal AgeI site found at 1808 - 1000COMPATIBLE 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.