Coding
Part:BBa_K5206005
Designed by: Yaoyao Liang Group: iGEM24_Nanjing-BioX (2024-09-30)
alsRm, alsR mutant
The BBa_K5206001 alsR gene from the genome of Escherichia coli BL21 (DE3) was subjected to targeted evolution by random mutagenesis with a view to obtaining mutants with improved functions.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Results
To construct alsR random mutant libraries, 25 μL of Mutazym, 2 μL of upstream and downstream primers (alsr - rtaq - FOR and alsR - rtaq - REV), different concentrations of manganese chloride (0.3-0.5 mM), and 10 ng of wild-type template DNA were added to the error-prone PCR reaction system, and the system was made up by adding ddH2O water.
Electrophoretic detection of PCR results
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Categories
Parameters
None |