Difference between revisions of "Part:BBa K4759229"

Revision as of 16:44, 10 October 2023

T7-RBS1-petH-RBS2-petF(D58F)

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

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 1336
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1336
Illegal NotI site found at 1109
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 1336
Illegal BglII site found at 1647
Illegal BamHI site found at 1330
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 1336
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 1336
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4759229 short</partinfo>
-<!-- Add more about the biology of this part here
 ===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%.
 <!-- -->