Difference between revisions of "Part:BBa K4937022"
18380928174 (Talk | contribs) |
18380928174 (Talk | contribs) |
||
| Line 5: | Line 5: | ||
<p>TDH3p-AtACL5-DIT1t:</p> | <p>TDH3p-AtACL5-DIT1t:</p> | ||
<p>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.</p> | <p>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.</p> | ||
| − | + | https://static.igem.wiki/teams/4937/wiki/part/021-1.png | |
| + | |||
<p>Using the OAZ1 knockout strain we previously constructed, we generated yeast expression plasmids to overexpress <i>At</i>ACL5, 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-<i>At</i>ACL5-DIT1t). Subsequently, we used homologous recombination to construct the complete plasmids and validated their successful construction through sequencing (Figure 2).</p> | <p>Using the OAZ1 knockout strain we previously constructed, we generated yeast expression plasmids to overexpress <i>At</i>ACL5, 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-<i>At</i>ACL5-DIT1t). Subsequently, we used homologous recombination to construct the complete plasmids and validated their successful construction through sequencing (Figure 2).</p> | ||
| − | <p> | + | https://static.igem.wiki/teams/4937/wiki/part/021-2.png |
| − | <p> | + | <p style=" text-align: center;">Figure 1</p> |
| + | https://static.igem.wiki/teams/4937/wiki/part/021-3.png | ||
| + | <p style=" text-align: center;">Figure 2</p> | ||
<p>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.</p> | <p>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.</p> | ||
| − | <p> | + | https://static.igem.wiki/teams/4937/wiki/part/021-4.png |
| − | + | <p style=" text-align: center;">Figure 3</p> | |
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
Revision as of 08:16, 28 September 2023
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.
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).
Figure 1
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.
Figure 3
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]