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:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE 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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