Difference between revisions of "Part:BBa K4119001"
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<partinfo>BBa_K4119001 short</partinfo> | <partinfo>BBa_K4119001 short</partinfo> | ||
| − | + | Hoping to make a useful contribution for future iGEM teams,we completed the experimental characterization of existing part, tsPurple, purple chromoprotein(incl RBS)(BBa_K1033905) and provided new documents for this part.<Br> | |
| + | Our team was very interested in the characterization of this chromoprotein and carried out the design and implementation of related experiments. | ||
| + | <Br> | ||
| + | In order to test the function of these chromoproteins, we constructed Pet-29a(+)-tsPurple. | ||
| + | <Br> | ||
| + | The expression was induced by T7 RNA polymerase provided by host cells. E.coil BL21 was used as receptor bacteria to express the protein, and then the fusion protein with histidine was purified by Nickle-affinity chromatography. If this chromoprotein are functional, we can observe the correct band through plasmid PCR, agarose gel electrophoresis, and the obvious color of the colony. After a series of purification operations, the correct band was observed by SDS-PAGE. Finally, use Mass Spectrometer to detect the exact molecular weight of the protein for final confirmation. | ||
| + | Our experiment results matched the general expected trend and data. | ||
| + | |||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
Revision as of 15:34, 12 October 2022
T7 promoter-lacO-tsPurple-6xHisTag-T7 terminator
Hoping to make a useful contribution for future iGEM teams,we completed the experimental characterization of existing part, tsPurple, purple chromoprotein(incl RBS)(BBa_K1033905) and provided new documents for this part.
Our team was very interested in the characterization of this chromoprotein and carried out the design and implementation of related experiments.
In order to test the function of these chromoproteins, we constructed Pet-29a(+)-tsPurple.
The expression was induced by T7 RNA polymerase provided by host cells. E.coil BL21 was used as receptor bacteria to express the protein, and then the fusion protein with histidine was purified by Nickle-affinity chromatography. If this chromoprotein are functional, we can observe the correct band through plasmid PCR, agarose gel electrophoresis, and the obvious color of the colony. After a series of purification operations, the correct band was observed by SDS-PAGE. Finally, use Mass Spectrometer to detect the exact molecular weight of the protein for final confirmation.
Our experiment results matched the general expected trend and data.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]