Difference between revisions of "Part:BBa K3002200:Experience"
(→User Reviews) |
|||
| Line 7: | Line 7: | ||
===User Reviews=== | ===User Reviews=== | ||
| + | <html> | ||
| + | <p> | ||
| + | Both MUT-PETase and MHETase were expressed in the cytosol of C.reinhardtii. The Mut-PETase is essential for the degradation of PET into MHET and MHETase for the degradation of MHET into terephthalic acid and ethylene glycol. The expression level of both enzymes in the cytosol seemed to be high. | ||
| + | |||
| + | |||
| + | |||
| + | <p></p><div class="figure"> | ||
| + | <img src="https://2019.igem.org/wiki/images/1/1b/T--TU_Kaiserslautern--resultsFigure4.svg"/> | ||
| + | <p class="caption"><span class="phat">Expression of the enzymes MUT-PETase and MHETase in <i>Chlamydomonas</i> <i>reinhardtii</i>. | ||
| + | </span><span class="accent">(a)</span> Level 2 MoClo construct harboring the aadA selection marker and the coding sequences for MUT-PETase, and MHETase (see Figure 1 for part description). <span class="accent">(b)</span> The UVM4 strain was transformed with the construct shown in <span class="accent">(a)</span>. 11 spectinomycin-resistant transformants were inoculated in TAP and samples taken after 3 days. Extracted whole-cell proteins were analysed by SDS-PAGE and immunoblotting using an anti-HA antibody. MW – molecular weight. The black arrow represents the MHETase, the white arrow the MUT-PETase. The expression of both MHETase (~70 kDa) and MUT-PETase (~35 kDa) is visible in colonies 18, 22 and 27. The UVM4 recipient strain and a strain expressing the HA-tagged ribosomal chloroplastic 50S protein L5 (RPL5) served as negative and positive controls, respectively. | ||
| + | </p> | ||
| + | </div><p> | ||
| + | |||
| + | |||
| + | |||
| + | </p> | ||
| + | </html> | ||
| + | |||
<!-- DON'T DELETE --><partinfo>BBa_K3002200 StartReviews</partinfo> | <!-- DON'T DELETE --><partinfo>BBa_K3002200 StartReviews</partinfo> | ||
<!-- Template for a user review | <!-- Template for a user review | ||
Latest revision as of 03:21, 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_K3002200
User Reviews
Both MUT-PETase and MHETase were expressed in the cytosol of C.reinhardtii. The Mut-PETase is essential for the degradation of PET into MHET and MHETase for the degradation of MHET into terephthalic acid and ethylene glycol. The expression level of both enzymes in the cytosol seemed to be high.
Expression of the enzymes MUT-PETase and MHETase in Chlamydomonas reinhardtii. (a) Level 2 MoClo construct harboring the aadA selection marker and the coding sequences for MUT-PETase, and MHETase (see Figure 1 for part description). (b) The UVM4 strain was transformed with the construct shown in (a). 11 spectinomycin-resistant transformants were inoculated in TAP and samples taken after 3 days. Extracted whole-cell proteins were analysed by SDS-PAGE and immunoblotting using an anti-HA antibody. MW – molecular weight. The black arrow represents the MHETase, the white arrow the MUT-PETase. The expression of both MHETase (~70 kDa) and MUT-PETase (~35 kDa) is visible in colonies 18, 22 and 27. The UVM4 recipient strain and a strain expressing the HA-tagged ribosomal chloroplastic 50S protein L5 (RPL5) served as negative and positive controls, respectively.
UNIQb07a71eb4c4981cf-partinfo-00000001-QINU UNIQb07a71eb4c4981cf-partinfo-00000002-QINU