Coding

Part:BBa_K2812006

Designed by: Guido Oerlemans, Maxime van den Oetelaar and Mariska Brüls   Group: iGEM18_TU-Eindhoven   (2018-10-03)
Revision as of 09:55, 15 October 2018 by MariskaBruls (Talk | contribs)


Coding sequence for Pyocin S5 with HlyA and His6-tag

Agarose gel of the pyocin S5 with HlyA and His6-tag.

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. 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.



[edit]
Categories
//cds
//cds/enzyme
//cds/enzyme/lysis
//cds/membrane
//cds/membrane/lysis
//chassis/prokaryote/ecoli
//collections/biofilm
//plasmid/expression
//plasmidbackbone/proteinfusion
//plasmidbackbone/synthesis
//proteindomain
//proteindomain/cleavage
//proteindomain/degradation
//proteindomain/linker
//proteindomain/localization
Parameters
None