View source for Part:BBa K4361303


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<partinfo>BBa_K4361303 short</partinfo>

A mutant of the BlcR protein, created through site-directed mutagenesis with primers R7 ([[Part:BBa_K4361210]]) and F7.1 S61V ([[Part:BBa_K4361211]]). For this mutant, the serine in position 61 has been changed to valine by mutating the AGT codon to GTG.

This mutant also contains the following nucleotide mutations outside of the targeted site:
<ul>

<li>C 705 > T, silent mutation</li>
</ul>

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===Usage and Biology===

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<span class='h3bb'><h3>Sequence and Features</h3></span>
<partinfo>BBa_K4361303 SequenceAndFeatures</partinfo>

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<h3>Usage and Biology</h3>
The set of BlcR mutants (</html>[[Part:BBa_K4361300]] through [[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 4</u>), a protein band corresponding to the BlcR monomer is visible. Concluding that the production in an E. coli cell-free system was successful for this mutant.

<figure>
<a href="https://static.igem.wiki/teams/4361/wiki/results/figure-7-pure/figure-7-pure-4.png"><img src="https://static.igem.wiki/teams/4361/wiki/results/figure-7-pure/figure-7-pure-4.png" style="width:600px;margin-left:125px"></a>
<figcaption> <b>Figure 1.</b> SDS PAGE showing the translation products of 20 different plasmids coding for BlcR mutants. PURE reaction solution was supplemented with GreenLys and the synthesized proteins were imaged with Typhoon using a 488-nm laser. C: negative control with no plasmid. WT: positive control with expressed wildtype BlcR. 4: S61V mutant</figcaption>
</figure>


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===Functional Parameters===
<partinfo>BBa_K4361303 parameters</partinfo>
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