Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Josh Atkinson, Mike Ferguson, and Ben Parker
Date created: 2012-09-28 11:00:00
Date modified: 2015-05-08 01:13:40
HbiF Recombinase (RFC25)
| Types | DnaRegion |
| Roles | Protein_Domain polypeptide_domain |
| Sequences | BBa_K880000_sequence (Version 1) |
Description
-HbiF-RFC25The Hbif recombinase is a putative regulator of the Escherichia coli type 1 fimbriation system, capable of switching the fimS invertible region in the production of fimbriae in a manner similar to the better characterized FimB and FimE recombinases. Orientation control of the fimS switch by recombinases is specific to directionality, with Hbif catalysing the conversion of ???OFF??? oriented regions to ???ON??? oriented ones such that the fimS promoter is facing genes responsible for type 1 fimbriae production.
The hbiF gene was synthesized in vitro as device capable of opposing the directional inversion of the characterized fimE gene K137007 for the purpose of engineering bidirectional molecular switches. A synthetic circuit can be made by fusing two unique invertable repeats (IRs) recognized by the recombinase-parts K137008 and K137010, in that order- flanking a sequence of interest of 200-300bp and expressing hbiF or fimE in low-to-moderate levels to cause the sequence to invert:
fimE will cause a template strand between the IRs to face the upstream coding strand.
hbiF will cause a coding strand between the IRs to face the downstream template strand.
Note that IRs??? initial orientation is FimE flippable, but not HbiF flippable.
This reaction can be characterized by digesting asymmetrical digest reporter K880001 with EcoRI and AgeI and determining the lengths of the resulting fragments.
Additionally, HbiF contains an RFC25 suffix and is capable of undergoing protein fusions to fluorescent proteins for quantification, affinity purification tags, or degradation tags such as K880004 to increase circuit responsiveness.
Reference: Xie et al. Infect. Immun. 2006, 74(7):4039