Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Date created: 2010-10-06 11:00:00
Date modified: 2015-05-08 01:11:55
| Types | DnaRegion |
| Roles | Temporary sequence_feature |
| Sequences | BBa_K314982_sequence (Version 1) |
Description
Our overall goal is to clone the Type 6 secretion system and the Tse2/Tsi2 toxin/antitoxin system from Pseudomonas aeruginosa into E. coli to make a strain of E. coli that could be used to kill off gram negative pathogens present in the human gut. Ideally, this system would be regulated in such a way that the strain of E. coli would only be able to kill gram negative bacteria when a gram negative pathogen is present. This strain could (ideally) be introduced into the gut either as a preventive measure or as a treatment after a known infection. Ideally, the probiotic would only kill of gram negative bacteria in the area of infection.Notes
All the essential genes for our T6SS are contained within two putative operons, encoded in opposite directions. The native promoters for both operons are found in the same intergenic region, between fha1 and tssA1. Therefore, we can easily replace the promoter regions for both operons in one step.Source
In order to create a probiotic application for this system, we first attempt to express it heterologously in non-pathogenic E. Coli. Starting from a Fosmid containing our T6SS, we are using Recombineering to replace the strict native regulation with robust T7 promoters to create strong expression of the T6SS.| Sequence Annotation | Location | Component / Role(s) |
| Promoter one Promoter 2 rbs StOP Tse2 | 1,17 12,35 12,67 56,78 50,75 | feature/promoter promoter promoter feature/promoter feature/rbs ribosome_entry_site feature/stop stop_codon feature/cds CDS |