Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Daniela Mehech
Date created: 2010-10-26 11:00:00
Date modified: 2015-05-08 01:12:29
Self-Lysis Device under Pbad and with BRP
| Types | DnaRegion |
| Roles | Composite engineered_region |
| Sequences | BBa_K426020_sequence (Version 1) |
Description
The Self Lysis Device acts to degrade the bacterial membrane (inner membrane, peptidoglycan layer and outer membrane).The Construct
Pbad promoter: allows for fast, controlled induction, allows better control of the timing of lysis, addresses biosafety concerns since this promoter is only induced by an exogenous molecule.
Bacteriocin Releasing Protein (BRP): degrades the outer membrane
Lysozyme: degrades the peptidoglycan layer
Holin: creates pores in the inner membrane
Anti-holin: threshold-gating system
(for more complete descriptions, see 2008 UC Berkeley iGEM team's wiki)
A variation of this part (that does not include BRP) was characterized in standard E. coli media, Luria Broth (LB), by the 2008 UC Berkeley iGEM team. In the Anderson lab, the construct was modified to include the BRP protein and has been extensively tested as a means of delivery in Mammalian cells. The 2010 Berkeley iGEM team, characterized the device in an entirely new setting: for use inside of Choanoflagellates.
Self-lysis can be assayed by measuring changes in optical density, or changes in the opaqueness of a solution. As lysis occurs, optical density decreases, meaning the solution of bacteria becomes more translucent. A TECAN machine was used to measure the change in optical density over time. The self-lysis device was tested in LB, TB, artificial sea water (ASW), and Cereal Grass Media(CGM). Tests showed that TB is the best media for lysis, followed by LB, CGM, high ratios of LB:ASW, low ratios of LB: ASW, and ASW. Also, macroscopic tests indicate that lysis occurs in spring water:LB ratios as high as 80:20.
Notes
NASource
SyntheticReferences:
iGEM 2010, UC Berkeley: http://2010.igem.org/Team:Berkeley/Project/Self_Lysis
iGEM 2008, UC Berkeley: http://2008.igem.org/Team:UC_Berkeley/LysisDevice
| Sequence Annotation | Location | Component / Role(s) |
| Pbad rbs.lambda lysozyme rbs.lambda holin rbs.antiholin Pcon rbs.BRP dbl Terminator rnpB T1 Terminator | 1,1194 1418,1919 1920,2263 2440,2789 2404,2438 1245,1417 2269,2397 2796,2908 | feature/promoter promoter CDS feature/protein CDS feature/protein CDS feature/protein promoter feature/promoter feature/protein CDS stem_loop feature/stem_loop feature/stem_loop stem_loop |