Difference between revisions of "Part:BBa K314203"

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<partinfo>BBa_K314203 short</partinfo>
 
<partinfo>BBa_K314203 short</partinfo>
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<br> This part contains  the [https://parts.igem.org/wiki/index.php?title=Part:BBa_K314202 Tse2/Tsi2 locus] from ''Pseudomonas aeruginosa'' downstream from an HSL inducible promoter([https://parts.igem.org/wiki/index.php?title=Part:BBa_F2620 F2620])
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Tse2/Tsi2 w/HSL inducible promoter
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This part contains  the [https://parts.igem.org/wiki/index.php?title=Part:BBa_K314200 Tse2/Tsi2 locus] from ''Pseudomonas aeruginosa'' downstream from an HSL inducible promoter( [https://parts.igem.org/wiki/index.php?title=Part:BBa_F2620 F2620])
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===Usage and Biology===
 
===Usage and Biology===
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[https://parts.igem.org/wiki/index.php?title=Part:BBa_K314200 Tse2] is a toxin that can be exported into gram (-) bacteria by the Type VI secretion system, killing all gram (-) cells not expressing Tsi2 ( the antitoxin of Tse2). Since Tse2 is being coexpressed with Tsi2, cells transformed with this part will not  die.  Therefore, this part is useful for regulating secretion of Tse2.
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This composite part produces the toxin Tse2 and its antitoxin Tsi2 when cells are exposed to HSL.  [https://parts.igem.org/wiki/index.php?title=Part:BBa_K314200 Tse2] is a potent cytotoxin. [https://parts.igem.org/wiki/index.php?title=Part:BBa_K314201 Tsi2] is its antitoxin, which protects the host cell from death by Tse2. Tse2 is targeted for export from the host cell cytoplasm into the cytoplasm of Gram-negative bacteria by the Type VI secretion system, killing the target Gram-negative cells unless they also express Tsi2.  Therefore, this part is useful for regulating secretion of Tse2.
  
 
===Functional Parameters===  
 
===Functional Parameters===  
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This part has been tested in pSB3k3. ''E. coli'' transformed with this part showed Tse2 production in a broth culture containing HSL at a concentration of 10µM showed Tse2 expression, while no appreciable level of Tse2 was produced in an environment not containing HSL. This was verified by an anti-Tse2 western blot of cellular supernatant of colonies grown up in liquid cultures containing either 10µM HSL or no HSL for a period of approximatively 24 hours. [[Image:Washington_Tse2_Tsi2_Western.jpg|600px|center|thumb|''E. coli'' showed Tse2 production in response to HSL]]
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This part has been tested in pSB3K3. ''E. coli'' transformed with this part showed Tse2 production in a broth culture containing HSL at a concentration of 10µM, while no appreciable level of Tse2 was produced in an environment not containing HSL. This was verified by an anti-Tse2 western blot of cellular supernatant of colonies grown up in liquid cultures containing either 10µM HSL or no HSL for a period of approximatively 24 hours. [[Image:Washington_Tse2_Tsi2_Western.jpg|600px|center|thumb|''E. coli'' showed Tse2 production in response to HSL]]
 
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===Sequence and Features===
 
===Sequence and Features===
 
<partinfo>BBa_K314203 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K314203 SequenceAndFeatures</partinfo>
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====Note: This part does not contain the standard biobricks scar between Lux pR and B0034 and between B0034 and Tse2/Tsi2. Instead of the standard biobrick scar at these position, this part contains the Bglbrick scar GGATCT.====
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====Note: This part does not contain the standard biobricks scar between Lux pR and B0034 and between B0034 and Tse2/Tsi2. Instead of the standard BioBrick scar at these position, this part contains the [https://parts.igem.org/Assembly_standard_21 BglBrick] scar GGATCT.====

Latest revision as of 23:45, 21 October 2010

HSL inducible toxin antitoxin, Tse2 + Tsi2
This part contains the Tse2/Tsi2 locus from Pseudomonas aeruginosa downstream from an HSL inducible promoter(F2620)

Usage and Biology

This composite part produces the toxin Tse2 and its antitoxin Tsi2 when cells are exposed to HSL. Tse2 is a potent cytotoxin. Tsi2 is its antitoxin, which protects the host cell from death by Tse2. Tse2 is targeted for export from the host cell cytoplasm into the cytoplasm of Gram-negative bacteria by the Type VI secretion system, killing the target Gram-negative cells unless they also express Tsi2. Therefore, this part is useful for regulating secretion of Tse2.

Functional Parameters

This part has been tested in pSB3K3. E. coli transformed with this part showed Tse2 production in a broth culture containing HSL at a concentration of 10µM, while no appreciable level of Tse2 was produced in an environment not containing HSL. This was verified by an anti-Tse2 western blot of cellular supernatant of colonies grown up in liquid cultures containing either 10µM HSL or no HSL for a period of approximatively 24 hours.
E. coli showed Tse2 production in response to HSL

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1283
    Illegal NgoMIV site found at 1340
    Illegal NgoMIV site found at 1465
    Illegal NgoMIV site found at 1795
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1004
    Illegal SapI.rc site found at 1790

Note: This part does not contain the standard biobricks scar between Lux pR and B0034 and between B0034 and Tse2/Tsi2. Instead of the standard BioBrick scar at these position, this part contains the BglBrick scar GGATCT.