Difference between revisions of "Part:BBa K4759214"
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<partinfo>BBa_K4759214 short</partinfo> | <partinfo>BBa_K4759214 short</partinfo> | ||
| − | + | T7-RBS1-petH-RBS2-petF-linker-GFP1-10 is constructed to selected for the best conventional redox partners | |
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| + | ===Usage and Biology=== | ||
| + | Generally, the method of determining whether the redox partner is suitable is through tedious steps such as the construction of plasmids, heterologous expression, construction of catalytic systems, and detection of conversion rate after catalysis. Therefore, we wanted to find a convenient way to do a quick screening. We used the fluorescent protein sfGFP to successfully construct a sensor to detect redox partners. sfGFP is a superfolder fluorescent protein that emits green light when irradiated with ultraviolet light. What is special about it is that it can be broken into two parts. | ||
| + | We divide sfGFP into sfGFP-1-10 and sfGFP-11, and although these two parts are cut off, there is an interaction force between them, and as long as they are properly folded in space, they will emit light again. Thus, four iron redox proteins are fused to the N-terminus of sfGFP-1-10 and Olep to the C-terminus of sfGFP-11, respectively, to obtain the recombinant plasmid pRSFDuet-BM3-GFP-1-10-GFP-11-oleP, pRSFDuet-camA-camB-GFP-1-10-GFP-11-oleP, pRSFDuet-FdR_0978-Fdx_1499-GFP-1-10-GFP-11-oleP, pRSFDuet-petH-petF-GFP-1-10-GFP-11-oleP | ||
| + | The above four recombinant plasmids are converted to BL21(DE3) to obtain recombinant strains G2 strain to G5 strain. | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
Revision as of 16:20, 10 October 2023
T7-RBS1-petH-RBS2-petF-linker-GFP1-10
T7-RBS1-petH-RBS2-petF-linker-GFP1-10 is constructed to selected for the best conventional redox partners
Usage and Biology
Generally, the method of determining whether the redox partner is suitable is through tedious steps such as the construction of plasmids, heterologous expression, construction of catalytic systems, and detection of conversion rate after catalysis. Therefore, we wanted to find a convenient way to do a quick screening. We used the fluorescent protein sfGFP to successfully construct a sensor to detect redox partners. sfGFP is a superfolder fluorescent protein that emits green light when irradiated with ultraviolet light. What is special about it is that it can be broken into two parts. We divide sfGFP into sfGFP-1-10 and sfGFP-11, and although these two parts are cut off, there is an interaction force between them, and as long as they are properly folded in space, they will emit light again. Thus, four iron redox proteins are fused to the N-terminus of sfGFP-1-10 and Olep to the C-terminus of sfGFP-11, respectively, to obtain the recombinant plasmid pRSFDuet-BM3-GFP-1-10-GFP-11-oleP, pRSFDuet-camA-camB-GFP-1-10-GFP-11-oleP, pRSFDuet-FdR_0978-Fdx_1499-GFP-1-10-GFP-11-oleP, pRSFDuet-petH-petF-GFP-1-10-GFP-11-oleP The above four recombinant plasmids are converted to BL21(DE3) to obtain recombinant strains G2 strain to G5 strain. Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1336
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1336
Illegal NotI site found at 1109 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1336
Illegal BglII site found at 1647
Illegal BglII site found at 2316
Illegal BamHI site found at 1330 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1336
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1336
- 1000COMPATIBLE WITH RFC[1000]