Difference between revisions of "Part:BBa K2812007"

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<partinfo>BBa_K2812007 short</partinfo>
 
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[[File:BBa_K2812007_Gel_1kb_Arabinose+Pyosin.png|100px|right|Agarose gel of the pyocin S5 with HlyA and His6-tag under control of the pBAD promoter.|thumb]]
 
Coding sequence for Pyocin S5, which is a bacteriocin produced by a specific strain of P. aeruginosa and is effective against other strains of P. aeruginosa. In this biobrick, Pyocin S5 is under control of the pBAD (araPb) promoter (BBa_K2812007). The sequence encodes for an effector component that consists of a receptor binding domain, a translocation domain and a killing domain. Furthermore it contains an immunity component which gives immunity against the bacteriocin. The HlyA tag is a secretion signal peptide which is used for secretion via the Type 1 secretion pathway of gram-negative bacteria. In order to facilitate protein purification, an N-terminal HIS-tag is present.
 
Coding sequence for Pyocin S5, which is a bacteriocin produced by a specific strain of P. aeruginosa and is effective against other strains of P. aeruginosa. In this biobrick, Pyocin S5 is under control of the pBAD (araPb) promoter (BBa_K2812007). The sequence encodes for an effector component that consists of a receptor binding domain, a translocation domain and a killing domain. Furthermore it contains an immunity component which gives immunity against the bacteriocin. The HlyA tag is a secretion signal peptide which is used for secretion via the Type 1 secretion pathway of gram-negative bacteria. In order to facilitate protein purification, an N-terminal HIS-tag is present.
  

Revision as of 09:58, 15 October 2018


Coding sequence for Pyocin S5 with HlyA and His6-tag regulated by pBAD-ara promoter

Agarose gel of the pyocin S5 with HlyA and His6-tag under control of the pBAD promoter.

Coding sequence for Pyocin S5, which is a bacteriocin produced by a specific strain of P. aeruginosa and is effective against other strains of P. aeruginosa. In this biobrick, Pyocin S5 is under control of the pBAD (araPb) promoter (BBa_K2812007). The sequence encodes for an effector component that consists of a receptor binding domain, a translocation domain and a killing domain. Furthermore it contains an immunity component which gives immunity against the bacteriocin. The HlyA tag is a secretion signal peptide which is used for secretion via the Type 1 secretion pathway of gram-negative bacteria. In order to facilitate protein purification, an N-terminal HIS-tag is present.

Usage and Biology

The working mechanism of Pyocin S5 involves making the target cell membrane permeable, causing leakage of intracellular materials. It has been demonstrated that is has antimicrobial activity against seven P. aeruginosa strains (DWW3, In3, In4, In7, In8, InA, and InB). P. aeruginosa PAO1, E. coli (BL21 (DE3), W3110, and ATCC15597) and S. aureus ATCC6538 were insensitive to pyocin S5. We as iGem Eindhoven 2018 aimed to use this biobrick for prevention of wound infections.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 208
    Illegal BglII site found at 496
    Illegal BglII site found at 2181
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1271