Fla-Art175

BBa_K1659001 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K1659001
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Raphaella Hull
Date created: 2015-08-09 11:00:00
Date modified: 2016-02-01 12:51:52

Artilysin Art-175 fused at N-terminal with flagellin 26-47 peptide segment



Types
DnaRegion

Roles
engineered_region

Composite

Sequences BBa_K1659001_sequence (Version 1)

Description

Artilysins are an exciting class of enzyme-based antibacterials. Their name is derived from "artificial endolysin" and they exploit the lytic power of bacteriophage-encoded endolyins. Endolysins are peptidoglycan hydrolases produced at the end of the lytic cycle that pass through the cytoplasmic membrane, degrade the peptidoglycan layer and cause the osmotic lysis of the infected bacterial cell, thus liberating the progeny. Purified endolysins have been used to kill Gram-positive pathogens. Gram-negative bacteria, however, have a protective outer membrane containing lipopolysaccharide (LPS) that serves as a barrier against the peptidoglycan hydolytic activity of endolysins from the outside.

To overcome this problem, selected polycationic or amphipathic peptides that locally destabilize the LPS layer have been covalently fused to endolysins. Biers et al. fused the sheep myeloid antimicrobial peptide (SMAP-29) to the N terminus of the endolysin KZ144 to create Artilysin 175. Art-175 has been shown to be a highly potent antibacterial that acts in minutes to kill virtually all Pseudomonas aeruginosa strains. [1]

BBa_K1659001 is a composite of Art-175 with the 26-47 segment of Salmonella typhimurium flagellin combined with a hexahistidine tag. [2]
Fla 26-47 is capable of mediating flagellar export of recombinant proteins and, combined with a hexahistidine tag, facilitates their secreted expression and easy purification.

[1] Briers, Y., Walmagh, M., Grymonprez, B., Biebl, M., Pirnay, J. P., Defraine, V., ??? Lavigne, R. (2014). Art-175 is a highly efficient antibacterial against multidrug-resistant strains and persisters of Pseudomonas aeruginosa. Antimicrobial Agents and Chemotherapy, 58(7), 3774???3784. http://doi.org/10.1128/AAC.02668-14
[2] Balb??s, P., & Lorence, A. (2004). Recombinant Gene Expression. Reviews and Protocols, 267, 506. http://doi.org/10.1007/978-1-61779-433-9

Notes

Export signal combined with hexahistidine tag

Source

Art-175 amino acid sequence: Briers, Y., Walmagh, M., Grymonprez, B., Biebl, M., Pirnay, J. P., Defraine, V., ??? Lavigne, R. (2014). Art-175 is a highly efficient antibacterial against multidrug-resistant strains and persisters of Pseudomonas aeruginosa. Antimicrobial Agents and Chemotherapy, 58(7), 3774???3784. http://doi.org/10.1128/AAC.02668-14

Amino acid sequence of Fla 26-47: Balb??s, P., & Lorence, A. (2004). Recombinant Gene Expression. Reviews and Protocols, 267, 506. http://doi.org/10.1007/978-1-61779-433-9

Sequence Annotation Location Component / Role(s)
Flagellin 26-47 Secretion Signal Tag
start
Art-175
Hisx6
stop
stop
4,165
1,3
166,1032
1033,1050
1051,1053
1054,1056
feature/tag tag
start_codon feature/start
CDS feature/cds
tag feature/tag
feature/stop stop_codon
stop_codon feature/stop
igem#experience
None
 
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K1659001/1