Difference between revisions of "Part:BBa K4361300"
Line 20: Line 20:
 
<html>
 
<html>
 
<h3>Usage and Biology</h3>
 
<h3>Usage and Biology</h3>
−
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 succesful 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.
  
 
<figure>
 
<figure>

Revision as of 12:54, 11 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:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 694
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 78
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 589

Usage and Biology

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.
Figure 1. SDS PAGE gel of PURE reactions with GreenLys and a variant of Part:BBa_K4361106 containing a BlcR mutant. Arrows indicate bands corresponding to BlcR dimers and monomers, respectively. C = negative control, WT = wildtype BlcR.