Part:BBa_K4759230
T7-RBS1-petH-RBS2-petF(D61I)
Usage and Biology
After obtaining the best redox partner, we performed alanine scanning on petF to speculate which sites had a greater impact on its electron transport capacity. Finally, we found that after mutations in seven of them, the electron transport effect would change greatly, so we mutated the amino acids of these sites into other 19 amino acids by modeling and selected 23 of them to show better results. We conducted control tests with the positive control group, negative control group, and wild-type strains, and finally selected 9 mutants with the highest fluorescence intensity for subsequent catalytic verification by detecting their green fluorescence intensity. By verifying the catalytic ability, We found that the substrate conversion of D68P was higher than that of the wild type in the nine strains with high fluorescence intensity, reaching 89.2%.
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 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]
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