Difference between revisions of "Part:BBa K4785005:Design"
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===Design Notes=== | ===Design Notes=== | ||
− | + | In order to examine the functionality of the HMGB1 protein and its domains, the subsequent four plasmids were designed to carry out protein expression. | |
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− | <img src="https://static.igem.wiki/teams/4785/wiki/hmgb1-plasmid.png" width=" | + | <img src="https://static.igem.wiki/teams/4785/wiki/hmgb1-plasmid.png" width="500" height="auto"> |
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− | Figure 1: To purify the target proteins, we construct five plasmids: pET28a-6×His- | + | Figure 1: To purify the target proteins, we construct five plasmids: pET28a-6×His-HMGB1_FL, pET28a-6×His-HMGB1_AB, pET28a-6×His-HMGB1_A, pET28a-6×His-HMGB1_B. After sequencing, we determine that the plasmid was correctly sequenced and ready for use. |
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− | <img src="https://static.igem.wiki/teams/4785/wiki/ | + | <img src="https://static.igem.wiki/teams/4785/wiki/.png" width="500" height="auto"> |
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− | Figure 2: The plasmids with the correct sequencing results were transferred into Escherichia coli BL21 for induced expression, then Escherichia coli was broken, and the target protein was purified by affinity chromatography and molecular sieve, and we obtained high purity of the target protein. | + | Figure 2: The plasmids with the correct sequencing results were transferred into Escherichia coli BL21 for induced expression, then Escherichia coli was broken, and the target protein was purified by affinity chromatography and molecular sieve, and we obtained high purity of the target protein. (a), SDS-PAGE of HMGB1_A. (b), SDS-PAGE of HMGB1_B. (c), SDS-PAGE of HMGB1_AB. (d), SDS-PAGE of HMGB1_FL. |
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+ | HMGB1_A and HMGB1_B appear to be more highly expressed in E. coli, presumably due to their shortened sequence lengths and reduced number of modifications, as determined experimentally. | ||
===Source=== | ===Source=== |
Latest revision as of 03:03, 12 October 2023
high-mobility group box 1 (hmgb1)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 73
Design Notes
In order to examine the functionality of the HMGB1 protein and its domains, the subsequent four plasmids were designed to carry out protein expression.
![](https://static.igem.wiki/teams/4785/wiki/hmgb1-plasmid.png)
![](https://static.igem.wiki/teams/4785/wiki/.png)
HMGB1_A and HMGB1_B appear to be more highly expressed in E. coli, presumably due to their shortened sequence lengths and reduced number of modifications, as determined experimentally.
Source
Organism: Homo sapiens