BBa_K541000

BBa_K541000 Version 1

Component

This part has been discontinued.

Please consider using BBa_K143053 as a replacement.

Source:
http://parts.igem.org/Part:BBa_K541000
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
engineered_region

Generator

Sequences BBa_K541000_sequence (Version 1)

Description

Constitutive promoter veg(BBa_K143012) coupled to the strong Ribosome Binding Site spoVG(BBa_K143021) from B. subtilis. These parts have been 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 circulatory system, 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

Access Instance Definition
public
BBa_K143053
Pveg-spoVG
Sequence Annotation Location Component / Role(s)
BBa_K143053
1,117
Pveg-spoVG
igem#experience
None
 
igem#partStatus
Released HQ 2013
igem#sampleStatus
Discontinued
igem#status
Deleted
 
isReplacedBy
BBa_K143053/1
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K541000/1