Composite

Part:BBa_K4937022

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


TDH3p-AtACL5-DIT1t

TDH3p-AtACL5-DIT1t:

This composite part was created by fusion PCR and used to exogenously express AtACL5 gene and results in the overexpression of thermospermine. This part was used in the oaz1Δ strain, in which the polyamine synthesis flux is stronger. We investigate whether this part can confer thermo-tolerance to resulted strain.

021-1.png

Using the OAZ1 knockout strain we previously constructed, we generated yeast expression plasmids to overexpress AtACL5, to achieve overexpression of thermospermine. We initially obtained the plasmid backbone through PCR (Figure 1: 7-1 to 7-4) and the insert fragments (Figure 1: 7-6 is 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 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
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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