Difference between revisions of "Part:BBa K4621002"
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chi18B hypothetical promoter is a functional fragment in chitinase Chi18B gene, which was cloned from SCUT-3 gene. In our project, through transcriptome analysis, qPCR and other means, we found that the gene expression of chitinase Chi18B was up-regulated when SCUT-3 was in chitin medium, and we speculated that its promoter chi18B hypothetical promoter may have properties that can be induced by chitin.
 
chi18B hypothetical promoter is a functional fragment in chitinase Chi18B gene, which was cloned from SCUT-3 gene. In our project, through transcriptome analysis, qPCR and other means, we found that the gene expression of chitinase Chi18B was up-regulated when SCUT-3 was in chitin medium, and we speculated that its promoter chi18B hypothetical promoter may have properties that can be induced by chitin.
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Clonal electrophoresis images of the fragments are shown in Figure. 1, with lane 2 representing chi18B hypothetical promoter
 
Clonal electrophoresis images of the fragments are shown in Figure. 1, with lane 2 representing chi18B hypothetical promoter
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chi18B hypothetical promoter and genes encoding a series of enzymes responsible for 5-Hydroxyectoine synthesis (ectoperon, ectD.et) were overexpressed using an integrated plasmid in Streptomyces SCUT-3. The modified strain was inoculated in 40 mL LB medium and fermented at 40℃ and 220 rpm for 120h. The production of Ectoine and 5-Hydroxyectoin in the presence or without chitin (the first day of fermentation) was detected to evaluate the chitin inducability of the promoter. The results are shown in Figure. 2.
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https://static.igem.wiki/teams/4621/wiki/parts/gel-electrophoresis-results.png
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By adding chitin at different days after fermentation, we also determined that the optimal time for chitin induction of chi18B hypothetical promoter was the first day of fermentation, and the specific experimental data are shown in Figure 3:
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chi18B hypothetical promoter and genes encoding a series of enzymes responsible for 5-Hydroxyectoine synthesis (ectoperon) were overexpressed using an integrated plasmid in Streptomyces SCUT-3. The modified strain was inoculated in 40 mL LB medium and fermented at 40℃ and 220 rpm for 120h. The production of Ectoine and 5-Hydroxyectoin in the presence or without chitin (the first day of fermentation) was detected to evaluate the chitin inducability of the promoter. The results are shown in Figure. 2.
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https://static.igem.wiki/teams/4621/wiki/parts/effect-of-different-promoters-on-yield.png
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Revision as of 18:27, 4 October 2023


A whole functional sequence in front of the chitinase Chi18B gene, including its promoter.


Usage, Biology and Characterization：

chi18B hypothetical promoter is a functional fragment in chitinase Chi18B gene, which was cloned from SCUT-3 gene. In our project, through transcriptome analysis, qPCR and other means, we found that the gene expression of chitinase Chi18B was up-regulated when SCUT-3 was in chitin medium, and we speculated that its promoter chi18B hypothetical promoter may have properties that can be induced by chitin.

Clonal electrophoresis images of the fragments are shown in Figure. 1, with lane 2 representing chi18B hypothetical promoter gel-electrophoresis-results.png

chi18B hypothetical promoter and genes encoding a series of enzymes responsible for 5-Hydroxyectoine synthesis (ectoperon) were overexpressed using an integrated plasmid in Streptomyces SCUT-3. The modified strain was inoculated in 40 mL LB medium and fermented at 40℃ and 220 rpm for 120h. The production of Ectoine and 5-Hydroxyectoin in the presence or without chitin (the first day of fermentation) was detected to evaluate the chitin inducability of the promoter. The results are shown in Figure. 2. effect-of-different-promoters-on-yield.png


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]