Difference between revisions of "Part:BBa K3002108:Experience"

 
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Construct is secreted trough secretion signal ARS. The secretion signal ARS shows minor secretion of the protein without combination with SP20 tag. Construct containing the ARS secretion signal leads to high yield of the secreted protein in comparison to the other secretion signals.
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<img src="https://2019.igem.org/wiki/images/6/6c/T--TU_Kaiserslautern--resultsFigure6.svg"/>
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<p class="caption"><span class="phat">Analysis of the secretion of MUT-PETase with secretion signals cCA, GLE, and ARS.                                                       
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</span><span class="accent">(a)</span> Level 2 MoClo constructs harboring the aadA selection marker, and the coding sequences for MUT-PETase equipped with secretion signals from carbonic anhydrase (cCA), gamete lytic enzyme (GLE) and arylsulfatase (ARS). See Figure 1 for the description of other parts. <span class="accent">(b)</span> UVM4 transformants containing the constructs shown in <span class="accent">(a)</span> were grown in TAP medium for seven days. Cells were centrifuged and the supernatant lyophilized, resuspended in 2xSDS buffer and analyzed by SDS-PAGE and immunoblotting with an anti-HA antibody.Transformants C12 and A27 introduced in Figures 4 and 5, respectively, served as positive controls. The white arrow points to the MUT-PETase.
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Latest revision as of 03:42, 22 October 2019


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_K3002108

User Reviews

Construct is secreted trough secretion signal ARS. The secretion signal ARS shows minor secretion of the protein without combination with SP20 tag. Construct containing the ARS secretion signal leads to high yield of the secreted protein in comparison to the other secretion signals.

Analysis of the secretion of MUT-PETase with secretion signals cCA, GLE, and ARS. (a) Level 2 MoClo constructs harboring the aadA selection marker, and the coding sequences for MUT-PETase equipped with secretion signals from carbonic anhydrase (cCA), gamete lytic enzyme (GLE) and arylsulfatase (ARS). See Figure 1 for the description of other parts. (b) UVM4 transformants containing the constructs shown in (a) were grown in TAP medium for seven days. Cells were centrifuged and the supernatant lyophilized, resuspended in 2xSDS buffer and analyzed by SDS-PAGE and immunoblotting with an anti-HA antibody.Transformants C12 and A27 introduced in Figures 4 and 5, respectively, served as positive controls. The white arrow points to the MUT-PETase.

</p> UNIQ6ee6d4e77a708612-partinfo-00000001-QINU UNIQ6ee6d4e77a708612-partinfo-00000002-QINU