Difference between revisions of "Part:BBa K2632002"
 
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Gene sifA is a key gene located in Salmonella Salmonella pathogenicity island and has the function of maintaining the integrity of Salmonella-Containing Vacuole (SCV) and was reported that it expresses after infection. A fragment including sifA promoter region and its’s RBS region was amplified from Salmonella SL1344 genome.   
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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 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.
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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).
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<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 (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>.
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<h1>Notice</h1>
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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>
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<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., Beuzón, 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 (2003).  3.          Beuzon, C. R. Salmonella maintains the integrity of its intracellular vacuole through the action of SifA. EMBO J. 19, 3235–3249 (2000).                    4.          Malik-Kale, P. et al. Salmonella- at home in the host cell. Front. Microbiol.2, 1–9 (2011)          5.          Steele-Mortimer, O. The Salmonella-containing vacuole-Moving with the times. Curr. Opin. Microbiol. 11, 38–45 (2008)(2008).</p>
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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 <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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<!-- Add more about the biology of this part here
 
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Latest revision as of 22:52, 17 October 2018

Promoter sifA

Gene sifA locates in Salmonella pathogenicity island-2 (SPI-2) and has the function of maintaining the integrity of Salmonella-Containing Vacuole (SCV). Some studies reported that the expression of SifA requires an intracellular enviroment. In other words, sifA promoter expresses SifA only when salmonella enters the host cells1. A fragment including sifA promoter region and RBS region was obtained by PCR from the Salmonella SL1344 genome. SipD is required for bacterial internalization. ΔsipD 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 PsifA (Figure 1).
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 carries it's native RBS. You can attach a CDS to it.
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

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).

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]