Difference between revisions of "Part:BBa K4169030"
Ruixuan Zhu (Talk | contribs) (→Functional Parameters) |
Ruixuan Zhu (Talk | contribs) (→Functional Parameters) |
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===Functional Parameters=== | ===Functional Parameters=== | ||
− | To verify the functionality of our part, | + | To verify the functionality of our part, we had the upstream gene of the promoter replaced with red fluorescence,and then transferred our plasmid into Escherichia coli DH5α and measured the intensity of red fluorescence with a microplate reader. Unfortunately, our bacteria did not produce red fluorescence, and upon analysis, it was conjectured that the transcriptional loxP formed a strong secondary structure and could not translate the red fluorescence protein behind it. |
<partinfo>BBa_K4169030 parameters</partinfo> | <partinfo>BBa_K4169030 parameters</partinfo> | ||
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Revision as of 04:21, 12 October 2022
Cre-reverse loxP system:working when you need to express genes in the other direction
In the presence of theophylline, expression of subsequent Cre recombinase is induced.In addition, the promoter and RBS between the two loxP will be reversed, making it express genes in the other direction.
Usage and Biology
When our engineered bacteria feel theophylline, Cre expresses it, Cre recognizes the reverse repeat sequence at both ends of the loxP site and combines to form a dimer, which then binds to the dimer on another loxP site to form a tetramer. This way Cre mediates the reverse sequence reversal between loxPs.
Functional Parameters
To verify the functionality of our part, we had the upstream gene of the promoter replaced with red fluorescence,and then transferred our plasmid into Escherichia coli DH5α and measured the intensity of red fluorescence with a microplate reader. Unfortunately, our bacteria did not produce red fluorescence, and upon analysis, it was conjectured that the transcriptional loxP formed a strong secondary structure and could not translate the red fluorescence protein behind it.
Sequence and Features
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 1292
Illegal NheI site found at 1315 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 454
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 39
Illegal AgeI site found at 140 - 1000COMPATIBLE WITH RFC[1000]