Difference between revisions of "Part:BBa K4361300"
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The set of BlcR mutants (this part through </html>[[Part:BBa_K4361319]]<html>) were expressed in a PURE system in the presence of GreenLys (fluorescent Cy2-labeled lysine). Samples were then run on an SDS PAGE gel (<b>Figure 1.</b>) to determine whether or not the protein was produced. For this mutant (<u>lane 1</u>), it was concluded that the production was not successful due to the bands corresponding to BlcR being too poorly distinguishable. | The set of BlcR mutants (this part through </html>[[Part:BBa_K4361319]]<html>) were expressed in a PURE system in the presence of GreenLys (fluorescent Cy2-labeled lysine). Samples were then run on an SDS PAGE gel (<b>Figure 1.</b>) to determine whether or not the protein was produced. For this mutant (<u>lane 1</u>), it was concluded that the production was not successful due to the bands corresponding to BlcR being too poorly distinguishable. | ||
Revision as of 15:25, 12 October 2022
BlcR D37R
A mutant of the BlcR protein (Part:BBa_K4361100), created through site-directed mutagenesis with primers R1 (Part:BBa_K4361200) and F1.1 D37R (Part:BBa_K4361201). For this mutant, the aspartic acid in position 37 has been changed to arginine by mutating the GAC codon to CGC.
This mutant also contains the following nucleotide mutations outside of the targeted site:
- G 763 > A, resulting in substitution E255K
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
Experimental results
The set of BlcR mutants (this part 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 1), it was concluded that the production was not successful due to the bands corresponding to BlcR being too poorly distinguishable.