Team:NYMU-Taipei/Project/Attachment

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Contents

Motivation

  • E. coli is a bacterium that commonly lives in the intestines of people and animals.

Goal

  • When genetic E.coli senses the alternation of pH in intestine, the adhesion mechanism will be turned on in order to enhance the ability of attachment.
  • The enhanced adhesion will make a chance for genetic E.coli to have more time to do things that has been designed by the "Clearance of Urea, Phosphate, and Guanidine" groups.
  • To cooperate with the "Time Regulation" group to allow the genetic E.coli to detach from intestinal epithelia cells after a specified amount of time.

How to attach in intestine

  • We have two strategies to enhance attachment in intestine: Expression of membrane form of FimH and FliC.

NYMU Attachment pic.jpg

  • We will use the biobrick part!
    • [http://partsregistry.org/Part:BBa_J04500 BBa_J04500]. a composite part of a lac promoter (R0010) and a strong ribosome binding site (B0034).
    • [http://partsregistry.org/Part:BBa_J36835 BBa_J36835]. Lpp, the lipoprotein signal peptide.
    • [http://partsregistry.org/Part:BBa_J36837 BBa_J36837] or [http://partsregistry.org/Part:BBa_J36838 BBa_J36838]. OmpA, one (O1) or five (O5) transmembrane domains, respectively. Both have been shown to work (Earhart, 2000).
    • [http://partsregistry.org/Part:BBa_J36841 BBa_J36841] or [http://partsregistry.org/Part:BBa_J36843 BBa_J36843]. Streptavidin, either wild-type "SW" (Howarth, 2006), or single-chain dimeric "SD" (Aslan, 2005).

What is FimH?

NYMU 7600693f1.jpg

  • The FimH protein is the receptor recognizing element of type 1 fimbriae
  • Fimbriae
    • Function: attachment of bacteria to its host as in case of pathogenic bacteria salmonella typhimurium, Nisseria gonorrhoea, bordella pertussis.
    • Size and number: Found many in number (up to 1000 per cell) having 3 to 25 nm in diameter and 0.5 to 20 micrometer in length.
  • Type 1 fimbriae are surface organelles of Escherichia coli which mediate D-mannose-sensitive binding to different host surfaces.
  • A single type 1 fimbria is a 7-nm-wide and »1-mm-long surface polymer, The bulk of the structure is made up of »1000 copies of the major subunit protein, FimA, polymerized into a right-handed helical structure.
  • Small quantities of the minor components (FimF, FimG, and FimH) are also present.
  • The FimH protein is the receptor recognizing element of type 1 fimbriae.
  • Studies on Salmonella Typhimurium revealed that FimH adhesin is responsible for bacteria binding to HeLa, HEp-2 and mouse intestinal epithelial cells.

What is FliC?

NYMU Flagellum.png

  • FliC is an antigen of E.coli flagella.
  • It binds to mucin and ECM.

Circuit Design

  • Preliminary Design
    • Design 1NYMU Attachment.jpg
    • Design 2NYMU 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]

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