Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Cihan TAŞTAN
Date created: 2011-07-12 11:00:00
Date modified: 2015-05-08 01:12:38
Limulus anti-LPS factor (LALF) for Bacillus subtilis
| Types | DnaRegion |
| Roles | Device engineered_region |
| Sequences | BBa_K541515_sequence (Version 1) |
Description
Constitutive promoter veg(BBa_K143012) coupled to the strong Ribosome Binding Site spoVG(BBa_K143021) from B. subtilis. These parts havebeen taken from 2008_Imperial_Collage. Pveg is a constitutive promoter that constitutively expresses the P43 protein in B. subtilis.
SpoVG is an endogenous ribosome binding site from B. subtilis. The sequence of the spoVG ribosome binding site is AAAGGUGGUGA which is
complementary to the sequence UUUCCUCCACU from the 3' region of the 16s rRNA from B. subtilis.
SacB is a signal peptide used in the Sec-SRP (secretory signal recognition particle) pathway by B. subtilis. Signal peptides are
responsible for directing preproteins (secretory proteins with a signal peptide region attached) through an appropriate secretory
pathway. In the case of the Sec-SRP signal peptide, they direct preproteins from the cytoplasm into the growth medium.
Lipopolysaccharide (LPS), or endotoxin, is the major mediator of septic shock, a serious complication of Gram-negative bacterial
infections in humans. Molecules that bind LPS and neutralize its biological effects or enhance its clearance could have important
clinical applications. Limulus anti-LPS factor (LALF) binds LPS
tightly, and, in animal models, reduces mortality when administered before or after LPS challenge or bacterial infection. The wedge-
shaped molecule has a striking charge distribution and amphipathicity that suggest how it can insert into membranes. The binding site
for LPS probably involves an extended amphipathic loop, and it has been proposed that two mammalian LPS-binding proteins will have a
similar loop. The amphipathic loop structure may be used in the design of molecules with therapeutic properties against septic shock.
Notes
During our design, we have looked out that this anti-LPS factor gene should be transfered into gram positive bacteria (i.e. B.subtilis) because of absence of LPS in gram positive bacteria.Source
Horseshoe crabs (Limulus polyphemus and Tachypleus tridentatus) are ancient arachnids that possess a primitive circulatorysystem, the hemolymph, containing only one kind of cell, the hemocyte. Exposure of hemocytes to bacterial endotoxins [lipopolysaccharide (LPS)] results in the activation of an intracellular coagulation cascade (Iwanaga et al., 1986), a defense against microbial invasion
| Sequence Annotation | Location | Component / Role(s) |
| BBa_K143012 BBa_K143053 Sigma A-35 Sigma A -10 rbs BBa_K143021 SacB LALF | 1,97 1,117 63,68 86,91 106,117 106,117 124,219 226,537 | engineered_region feature/BioBrick engineered_region feature/BioBrick feature/promoter promoter promoter feature/promoter ribosome_entry_site feature/rbs engineered_region feature/BioBrick tag feature/tag CDS feature/protein |