Difference between revisions of "Part:BBa K3002001"

 
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<partinfo>BBa_K3002001 short</partinfo>
 
<partinfo>BBa_K3002001 short</partinfo>
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<img src="https://2019.igem.org/wiki/images/1/1b/T--TU_Kaiserslautern--resultsFigure4.svg"/>
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<p class="caption"><span class="phat">Expression of the enzymes MUT-PETase and MHETase in <i>Chlamydomonas</i> <i>reinhardtii</i>.             
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</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.
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This basic part contains a combination of the PSAD promoter and a 5'UTR (A1-B2) for Chlamydomonas reinhardtii and was built as a part of the Kaiser Collection. Combined with a CDS and a terminator, this level 0 construct leads to a high expression of a target protein.
 
This basic part contains a combination of the PSAD promoter and a 5'UTR (A1-B2) for Chlamydomonas reinhardtii and was built as a part of the Kaiser Collection. Combined with a CDS and a terminator, this level 0 construct leads to a high expression of a target protein.

Revision as of 14:06, 10 December 2019


PSAD promoter for Chlamydomonas reinhardtii (Phytobrick)

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.

This basic part contains a combination of the PSAD promoter and a 5'UTR (A1-B2) for Chlamydomonas reinhardtii and was built as a part of the Kaiser Collection. Combined with a CDS and a terminator, this level 0 construct leads to a high expression of a target protein.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]