Team:Brown/Project

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Latest revision as of 21:07, 27 October 2010

Light-Pattern Controlled Circuit

Abstract

Biological manufacturing of complex compounds often requires the synthesis of many intermediate products. Production of these intermediates is currently triggered by inefficient methods, such as chemical inputs (tetracycline, estrogen-analogs, arabinose, etc) or drastic changes to the cellular environment (pH, oxygen levels, temperature, etc). On an industrial scale, this chemical induction requires large quantities of reagents and extensive purification, while environmental induction requires conditions that can adversely affect cell vitality and yield. To this end, we are engineering an E. coli genetic circuit that can pass through four stable states of protein production triggered solely by ON/OFF patterns of light. With this production method, we can link multiple synthesis steps to a single, clean and rapidly scalable input.

E. Cargo

Abstract

We designed a modular Tat-Linker Biobrick that will allow for easy fusion of the Tat-PTD to any other biobricked proteins. Specifically, we aimed to fuse this Tat-Linker in RFC25 format to two bacterial transcription factors, LacI and AraC, with the intent of using the two Tat-TFs as a tool to induce transient gene expression in E. coli without the need to follow through with more time-consuming cell transformation protocols. To test these Tat-TFs, we designed corresponding reporter constructs. We saw a potential application in the portion of the Light-Pattern Controlled Circuit of our project, as well as in any other future genetic circuits that require part-by-part testing.

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 <br>
-== '''Project 1 - Light-Pattern Controlled Circuit''' ==
+== '''Light-Pattern Controlled Circuit''' ==
-=== Project Description ===
+'''Abstract'''
-The aim of this project is to create a cell that can go through a sequence of four distinct states in response to a timed pattern of light-on, light-off.
+Biological manufacturing of complex compounds often requires the synthesis of many intermediate products. Production of these intermediates is currently triggered by inefficient methods, such as chemical inputs (tetracycline, estrogen-analogs, arabinose, etc) or drastic changes to the cellular environment (pH, oxygen levels, temperature, etc). On an industrial scale, this chemical induction requires large quantities of reagents and extensive purification, while environmental induction requires conditions that can adversely affect cell vitality and yield. To this end, '''we are engineering an E. coli genetic circuit that can pass through four stable states of protein production triggered solely by ON/OFF patterns of light.''' With this production method, '''we can link multiple synthesis steps to a single, clean and rapidly scalable input.'''
+<html>
+<center>
+<a href="https://2010.igem.org/Team:Brown/Project/Light_pattern/Overview"><img src="https://static.igem.org/mediawiki/2010/thumb/c/c2/LRCpeektacropped.jpg/780px-LRCpeektacropped.jpg" width="500px"></a>
+</center>
+</html>
-This project makes use of parts from EPF Lusanne '09 (the LOVTAP light activated promoter), PKU '07/'09 (bistable switch, AND gate), and Missouri Western State University '08 (Hybrid promoter). Our goal is to show that it is possible and easy to create entirely new cell logic systems from parts that already exist in the registry.
+== E. Cargo ==
-See the model for this system here:[[Team:Brown/Modeling]]
+'''Abstract'''
-== '''Project 2 - TAT-PTD''' ==
+We designed a modular Tat-Linker Biobrick that will allow for easy fusion of the Tat-PTD to any other biobricked proteins. Specifically, we aimed to fuse this Tat-Linker in RFC25 format to two bacterial transcription factors, LacI and AraC, with the intent of using the two Tat-TFs as a tool to induce transient gene expression in <i>E. coli</i> without the need to follow through with more time-consuming cell transformation protocols. To test these Tat-TFs, we designed corresponding reporter constructs. We saw a potential application in the portion of the Light-Pattern Controlled Circuit of our project, as well as in any other future genetic circuits that require part-by-part testing.
+<html>
-=== Project Description ===
+<center>
+<a href="https://2010.igem.org/Team:Brown/Project/Ecargo/Overview"><img src="https://static.igem.org/mediawiki/2010/thumb/8/8d/ECargo.png/400px-ECargo.png" width="500px"></a>
-We have created a recombinant protein which fuses a Tat-protein transduction domain (Tat-ptd) with a synthetic single chain variable fragment (scFv) domain. The goals of this project are: 1) be able to stably express the fusion protein in E. Coli; 2) create a protein that will retain conformation and function in cytosolic conditions; 3) the intrabody should be able to bind with high specificity to the desired target protein; 4) the intrabody can translocate across mammalian cell and tissue barriers; and 5) include a method for easy fluorescence reporting.
+</center>
+</html>
-We also are in the process of creating a biobrick that will allow for easy fusion of the Tat-PTD to another biobricked proteins. We plan on characterizing this biobrick by combining it with transcription factors in the Registry and  transiently acting on inducible/repressible reporter constructs.