Team:UST-Beijing/Project

From 2011.igem.org

(Difference between revisions)
(→Part 2)
Line 64: Line 64:
=== Part 2 ===
=== Part 2 ===
-
 
+
PR，which is a abbreviation of proteorhodopsin, was discovered in 2000 through shortgun sequencing of seawater off the coast of California. It is a a kind of menbrane proteins that generates a proton motive force(pmf) that can be used for adenosine triphosphate (ATP) synthesis as a light-activated proton pump. In our experiment, we replaced the PR’s precusor sequence with that of cytochrome oxidase subunit 4 isoform 1, mitochondrial precursor (Homo sapiens). So PR can be fixed in the inner membrane of mitochrondria. Meanwhile, all the codons are optimized according to codon usage bias for Homo sspiens. Then, the gene of PR is constructed in pSG5 vector for eukaryotic expression.
-
 
+
-
 
+
-
 
+
=== The Experiments ===
=== The Experiments ===

Revision as of 08:02, 4 September 2011


This is a template page. READ THESE INSTRUCTIONS.
You are provided with this team page template with which to start the iGEM season. You may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki. You can find some examples HERE.
You MUST have a team description page, a project abstract, a complete project description, a lab notebook, and a safety page. PLEASE keep all of your pages within your teams namespace.


You can write a background of your team here. Give us a background of your team, the members, etc. Or tell us more about something of your choosing.
UST-Beijing logo.png

Gene H-transfer: bile acid receptor in E.coli & proteorhodpsin in mitochindrial inner membrane


In order to celebrate the power of gene H(orizontal)-transfer between pro- and eukaryotes, we constructed two fusion proteins and tested their function: (1). a synthetic bile acid receptor in E.coli using a mammalian nuclear receptor LXR. As proof-of-principle, the regulatory circuit in symbiotic bacteria could be harmoniously linked to metabolic pathway of their host. Potential application includes in situ synthesis of pharmaceuticals on-demand in the digestive tract. (2) a synthetic light-driven proton pump in human mitocondrial inner membrane using a bacterial proteorhodopsin. Preliminary testing demonstrated cellular sensitivity to light radiation. Application and utility relies on result of in-depth characterization of such system design.

Your team picture
Team Example


Home Team Official Team Profile Project Parts Submitted to the Registry Modeling Notebook Safety Attributions



Contents

We are engineering a bile acid sensor in E.coli and others 

 Using lacI DNA-binding domain and LXRbeta ligand-binding domain, we made an artificial bile acid receptor
which can regulate expression of target gene within a natural lacI operon. As proof of principle, we

demonstrate that regulation of bacteria gene expression by host eukaryocyte metabolites is achievable using chimeric nuclear receptors. Through directed molecular evolution, a harmonious signal network regulating metabolism of both prokaryocytes and their host eukaryocytes in the digestive tract is feasible.

Project Details

Part 2

PR，which is a abbreviation of proteorhodopsin, was discovered in 2000 through shortgun sequencing of seawater off the coast of California. It is a a kind of menbrane proteins that generates a proton motive force(pmf) that can be used for adenosine triphosphate (ATP) synthesis as a light-activated proton pump. In our experiment, we replaced the PR’s precusor sequence with that of cytochrome oxidase subunit 4 isoform 1, mitochondrial precursor (Homo sapiens). So PR can be fixed in the inner membrane of mitochrondria. Meanwhile, all the codons are optimized according to codon usage bias for Homo sspiens. Then, the gene of PR is constructed in pSG5 vector for eukaryotic expression.

The Experiments

Part 3

Results

Retrieved from "http://2011.igem.org/Team:UST-Beijing/Project"