Difference between revisions of "Part:BBa K3002113:Experience"
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The MHETase in combination with the secretion signal cCA showed constant secretion by C.reinhardtii. This applies for the MHETase alone but also for the MHETase in combination with other level 1 constructs of the PETase. Constructs including cCA lead to a high yield of the secreted MHETase. | The MHETase in combination with the secretion signal cCA showed constant secretion by C.reinhardtii. This applies for the MHETase alone but also for the MHETase in combination with other level 1 constructs of the PETase. Constructs including cCA lead to a high yield of the secreted MHETase. | ||
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| − | <div class="figure"> | + | <p></p><div class="figure"> |
<img src="https://2019.igem.org/wiki/images/b/b8/T--TU_Kaiserslautern--resultsFigure5.svg"/> | <img src="https://2019.igem.org/wiki/images/b/b8/T--TU_Kaiserslautern--resultsFigure5.svg"/> | ||
<p class="caption"><span class="phat">MUT-PETase destined for secretion gets stuck inside the cell. | <p class="caption"><span class="phat">MUT-PETase destined for secretion gets stuck inside the cell. | ||
Revision as of 01:49, 22 October 2019
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Applications of BBa_K3002113
User Reviews
The MHETase in combination with the secretion signal cCA showed constant secretion by C.reinhardtii. This applies for the MHETase alone but also for the MHETase in combination with other level 1 constructs of the PETase. Constructs including cCA lead to a high yield of the secreted MHETase.
MUT-PETase destined for secretion gets stuck inside the cell. (a) Level 2 MoClo construct harboring the aadA selection marker, and the coding sequences for MUT-PETase and MHETase genes. MUT-PETase and MHETase are equipped with the secretion signal from carbonic anhydrase (cCA). See Figure 1 for the description of other parts. (b) Seven days old cultures of transformants generated with the construct shown in (a) were centrifuged and proteins in the culture medium were precipitated by TCA and analysed by immunoblotting using an anti-HA antibody. The black arrow represents MHETase. (c) Whole-cell proteins of UVM4 cells transformed with construct L2C shown in (a) were analyzed by immuno-blotting using an anti-HA antibody. Transformant A27 generated with construct L2A (Figure 4a) and UVM4 were used as positive and negative controls, respectively. The white arrow indicates MUT-PETase. (d) Immunfluorescence analysis of transformants 17 and 27 using an anti-HA antibody. DAPI staining was also performed. UVM4 cells served as control.
UNIQe70fe2c8d43f0469-partinfo-00000001-QINU UNIQe70fe2c8d43f0469-partinfo-00000002-QINU