Difference between revisions of "Part:BBa K1159105:Experience"
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The Sao Carlos-Brazil Team has developed a codon-optimized version of this part, it can be accessed at https://parts.igem.org/Part:BBa_K3273013
 
The Sao Carlos-Brazil Team has developed a codon-optimized version of this part, it can be accessed at https://parts.igem.org/Part:BBa_K3273013
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<partinfo>BBa_K1159105 AddReview 3</partinfo>
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<I>CWChew</I>
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Successfully cloned part into S. cerevisiae and tested part by placing nuclease downstream of the galactose inducible promoter Gal1 (Figure 1). Kill switch efficacy was not optimal and had a 82% survival rate.
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Attempted to perform part improvement by adding a nuclear localization signal tag to the 5' end of the part sequence to improve the kill switch efficacy in part <partinfo>Ba_K3927015</partinfo>.
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https://2021.igem.org/wiki/images/8/8a/T--NUS_Singapore--results_tab3_3.png
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<i> Figure 1: Construct schematic of part assembled with a Gal1 promoter upstream to trigger galactose dependent endonuclease activity</i>
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https://2021.igem.org/wiki/images/3/38/T--NUS_Singapore--results_tab3_5.png
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<i> Figure 2: CFU assay results from pGNucA-H(BY4741) and pGNLSNucA-H(BY4741), ratio of colonies counted from plates with the nuclease induced to the colonies counted from the plates with nuclease uninduced. Nuclease with the NLS attached reduces the total number of live colonies on the plate. </i>
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<!-- DON'T DELETE --><partinfo>BBa_K1159105 StartReviews</partinfo>
 
<!-- DON'T DELETE --><partinfo>BBa_K1159105 StartReviews</partinfo>

Revision as of 06:34, 18 October 2021


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1159105

User Reviews

The Sao Carlos-Brazil Team has developed a codon-optimized version of this part, it can be accessed at https://parts.igem.org/Part:BBa_K3273013

•••

CWChew

Successfully cloned part into S. cerevisiae and tested part by placing nuclease downstream of the galactose inducible promoter Gal1 (Figure 1). Kill switch efficacy was not optimal and had a 82% survival rate.

Attempted to perform part improvement by adding a nuclear localization signal tag to the 5' end of the part sequence to improve the kill switch efficacy in part BBa_K3927015.

T--NUS_Singapore--results_tab3_3.png

Figure 1: Construct schematic of part assembled with a Gal1 promoter upstream to trigger galactose dependent endonuclease activity

T--NUS_Singapore--results_tab3_5.png

Figure 2: CFU assay results from pGNucA-H(BY4741) and pGNLSNucA-H(BY4741), ratio of colonies counted from plates with the nuclease induced to the colonies counted from the plates with nuclease uninduced. Nuclease with the NLS attached reduces the total number of live colonies on the plate.

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UNIQcc79759c94096ebf-partinfo-00000002-QINU UNIQcc79759c94096ebf-partinfo-00000003-QINU