Difference between revisions of "Part:BBa K4361311"
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<partinfo>BBa_K4361311 short</partinfo> | <partinfo>BBa_K4361311 short</partinfo> | ||
| − | A mutant of the BlcR protein, created through site-directed mutagenesis with primers R3 ([[Part:BBa_K4361218]]) and F3.2 | + | A mutant of the BlcR protein, created through site-directed mutagenesis with primers R3 ([[Part:BBa_K4361218]]) and F3.2 L66A ([[Part:BBa_K4361220]]). For this mutant, the leucine in position 66 has been changed to alanine by mutating the CTC codon to GCG. |
This mutant also contains the following nucleotide mutations outside of the targeted site: | This mutant also contains the following nucleotide mutations outside of the targeted site: | ||
Revision as of 13:20, 11 October 2022
BlcR L66A
A mutant of the BlcR protein, created through site-directed mutagenesis with primers R3 (Part:BBa_K4361218) and F3.2 L66A (Part:BBa_K4361220). For this mutant, the leucine in position 66 has been changed to alanine by mutating the CTC codon to GCG.
This mutant also contains the following nucleotide mutations outside of the targeted site:
- T 826 > A, resulting in possible loss of the stop codon
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 694
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 78
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 589
Usage and Biology
The set of BlcR mutants (Part:BBa_K4361300 through Part:BBa_K4361319) were expressed in a PURE system in the presence of GreenLys (fluorescent Cy2-labeled lysine). Samples were then run on an SDS PAGE gel (Figure 1.) to determine whether or not the protein was produced. For this mutant (lane 12), a band was visible on the gel at the expected position, meaning that the production was successful for this mutant.