Composite

Part:BBa_K4937023

Designed by: Chen Wang   Group: iGEM23_SCU-BSE-China   (2023-09-27)
Revision as of 08:16, 28 September 2023 by 18380928174 (Talk | contribs)


TEF1p-SPE1-PRM9t-TDH3p-AtACL5-DIT1t

TEF1p-SPE1-PRM9t-TDH3p-AtACL5-DIT1t:

021-1.png

Using the OAZ1 knockout strain we previously constructed, we generated yeast expression plasmids to overexpress SPE1 and AtACL5, to achieve overexpression of putrescine (and subsequently spermidine) and thermospermine. We initially obtained the plasmid backbone through PCR (Figure 1: 7-1 to 7-4) and the insert fragments (Figure 1: 7-7 is TEF1p-SPE1-PRM9t-TDH3p-AtACL5-DIT1t). Subsequently, we used homologous recombination to construct the complete plasmids and validated their successful construction through sequencing (Figure 2).

021-2.png

Figure 1

021-3.png

Figure 2

Then we tested the growth of these strains under 35°C conditions (Figure 3). The results indicate that the expression of the SPE1-AtACL5 gene did not have a significant impact on the yeast's thermo-tolerance capabilities.

021-4.png

Figure 3


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1871
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1607
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 167


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