Team:NYMU-Taipei/Project/Attachment
From 2008.igem.org
Contents |
Motivation
- E. coli is a bacterium that commonly lives in the intestines of people and animals.
Goal
- When genetic E.coli sense the alternation of pH in intestine, adhesion mechanism will be turn on in order to enhance the ability of attachment.
- Enhanced adhesion will make a chance for genetic E.coli to have more time to do events that we design in "Clearance of urea (phosphat, guanidine)" group.
- To cooperate with "Time Regulation" group, we expect genetic E.coli to detach from intestinal epithelia cells at indicated time.
How to attach in intestine
- We have two strategies to enhance attachment in intestine: Expression of membrance form of FimH and FliC.
- We will use the biobrick part!
- BBa_J04500. a composite part of a lac promoter (R0010) and a strong ribosome binding site (B0034)[http://partsregistry.org/Part:BBa_J04500]
- BBa_J36835. Lpp, the lipoprotein signal peptide.[http://partsregistry.org/Part:BBa_J36835]
- BBa_J36837 or BBa_J36838. OmpA, one (O1) or five (O5) transmembrane domains, respectively. Both have been shown to work (Earhart, 2000).[http://partsregistry.org/wiki/index.php/Part:BBa_J36837][http://partsregistry.org/wiki/index.php/Part:BBa_J36838]
- BBa_J36841 or BBa_J36843. Streptavidin, either wild-type "SW" (Howarth, 2006), or single-chain dimeric "SD" (Aslan, 2005).[http://partsregistry.org/wiki/index.php/Part:BBa_J36841][http://partsregistry.org/wiki/index.php/Part:BBa_J36843]
What is FimH??
|
What is FliC??
|
Circuit Design
- Preliminary Design
- Design 1File:Attachment.jpg
- Design 2File:Combine 3.jpg
References
- Functional characterization of the FimH adhesin from Salmonella enterica serovar Enteritidis[http://mic.sgmjournals.org/cgi/content/full/152/5/1337?view=long&pmid=16622051]
- Combining sites of bacterial fimbriae [http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VS6-4PGPKS8-2&_user=1576506&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000053839&_version=1&_urlVersion=0&_userid=1576506&md5=33dfc6974f43b7ece79e0e7b90044ee7]
- Probing the receptor recognition site of the FimH adhesin by fimbriae- displayed FimH-FocH hybrids[http://mic.sgmjournals.org/cgi/reprint/144/7/1919?view=reprint&pmid=9695925]
- Host Protein Binding and Adhesive Properties of H6 and H7 Flagella of Attaching and Effacing Escherichia coli[http://jb.asm.org/cgi/content/full/189/20/7426]
- Sequence Diversity of the Escherichia coli H7 fliC Genes:Implication for a DNA-Based Typing Scheme forE. coli O157:H7[http://jcm.asm.org/cgi/reprint/38/5/1786.pdf]