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 was reported that expression of SifA is required for intracellular enviroment. In other words, promoter of <i>sifA</i> expresses SifA only when salmonella enter the host cells <sup>1</sup>. A fragment including sifA promoter region and RBS region were amplified from Salmonella SL1344 genome.
 
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 was reported that expression of SifA is required for intracellular enviroment. In other words, promoter of <i>sifA</i> expresses SifA only when salmonella enter the host cells <sup>1</sup>. A fragment including sifA promoter region and RBS region were amplified from Salmonella SL1344 genome.
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SipD is required for bacterial internalization. ΔsipD mutant can not enter host cells. Thus, we use this strain as a control. Microscopy suggests that only intracellular bacteria express eGFP which under the control of P<sub style:"font-family: italic">sifA</sub> (Fig.1).  
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SipD is required for bacterial internalization. ΔsipD mutant can not enter host cells. Thus, we use this strain as a control. Microscopy suggests that only intracellular bacteria express eGFP which 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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Revision as of 12:23, 13 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 was reported that expression of SifA is required for intracellular enviroment. In other words, promoter of sifA expresses SifA only when salmonella enter the host cells 1. A fragment including sifA promoter region and RBS region were amplified from Salmonella SL1344 genome. SipD is required for bacterial internalization. ΔsipD mutant can not enter host cells. Thus, we use this strain as a control. Microscopy suggests that only intracellular bacteria express eGFP which under the control of PsifA (Figure 1).
Figure 1. Hela GSDMD KO cells respectively infected with the ΔsipD or ΔsifA mutant. These mutants contain a low copy number plasmid to express eGFP which was regulated by PsifA.

Notice

This part carried 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 cell.

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]