Difference between revisions of "Part:BBa K2632002"
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− | Gene <i>sifA</i> locates in <i>Salmonella</i> pathogenicity island-2 (SPI-2) and has the function of maintaining the integrity of <i>Salmonella</i>-Containing Vacuole (SCV). Some studies | + | Gene <i>sifA</i> locates in <i>Salmonella</i> pathogenicity island-2 (SPI-2) and has the function of maintaining the integrity of <i>Salmonella</i>-Containing Vacuole (SCV). Some studies reported that the expression of SifA requires an intracellular enviroment. In other words, <i>sifA</i> promoter expresses SifA only when <i>salmonella</i> enters the host cells<sup>1</sup>. A fragment including sifA promoter region and RBS region was obtained by PCR from the <i>Salmonella</i> SL1344 genome. |
− | SipD is required for bacterial internalization. ΔsipD mutant | + | SipD is required for bacterial internalization. <i>ΔsipD</i> mutant cannot enter host cells. Thus, we use this strain as a control. Microscopy suggests that only intracellular bacteria express eGFP which is under the control of P<sub style:"font-family: italic">sifA</sub> (Figure 1). |
<img style="width: 800px; margin-left: 50px" src="https://static.igem.org/mediawiki/2018/9/90/T--HZAU-China--PsifA.jpg"> | <img style="width: 800px; margin-left: 50px" src="https://static.igem.org/mediawiki/2018/9/90/T--HZAU-China--PsifA.jpg"> | ||
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− | Figure 1. Hela GSDMD KO cells | + | Figure 1. Hela GSDMD KO (knockout) cells are infected with the <i>ΔsipD</i> or <i>ΔsifA</i> mutants, respectively. These mutants contain a low copy number plasmid to express eGFP which is regulated by P<sub style:"font-family: italic">sifA</sub>. |
<h1>Notice</h1> | <h1>Notice</h1> | ||
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− | This part | + | This part carries it's native RBS. You can attach a CDS to it. <br>This part should be in a low copy plasmid such as PSB4K5, or it will express before <i>Salmonella</i>'s entry to host cells. </br> |
− | <br> Use in <i>Salmonella</i> only.</br> | + | <br> <b>Use in <i>Salmonella</i> only.</b></br> |
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<h3>Reference</h3> | <h3>Reference</h3> | ||
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− | 1. Garmendia, J., Beuzon, C. R., Ruiz-Albert, J. & Holden, D. W. The roles of SsrA-SsrB and OmpR-EnvZ in the regulation of genes encoding the Salmonella typhimurium SPI-2 type III secretion system. Microbiology 149, 2385-2396, doi:10.1099/mic.0.26397-0 (2003). | + | 1. Garmendia, J., Beuzon, C. R., Ruiz-Albert, J. & Holden, D. W. The roles of SsrA-SsrB and OmpR-EnvZ in the regulation of genes encoding the <i>Salmonella</i> typhimurium SPI-2 type III secretion system. Microbiology 149, 2385-2396, doi:10.1099/mic.0.26397-0 (2003). |
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Latest revision as of 22:52, 17 October 2018
Promoter sifA
1. Garmendia, J., Beuzon, C. R., Ruiz-Albert, J. & Holden, D. W. The roles of SsrA-SsrB and OmpR-EnvZ in the regulation of genes encoding the Salmonella typhimurium SPI-2 type III secretion system. Microbiology 149, 2385-2396, doi:10.1099/mic.0.26397-0 (2003).
Figure 1. Hela GSDMD KO (knockout) cells are infected with the ΔsipD or ΔsifA mutants, respectively. These mutants contain a low copy number plasmid to express eGFP which is regulated by PsifA.
Notice
This part should be in a low copy plasmid such as PSB4K5, or it will express before Salmonella's entry to host cells.
Use in Salmonella only.
Reference
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]