Team:Penn State/PromoterProject

From 2013.igem.org

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             <h2 style="color: green" ID="Intro"> Introduction</h2>
             <h2 style="color: green" ID="Intro"> Introduction</h2>
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        Promoters are the one of the basic units of a genetic circuit. They tell an organism where genes start within the genome. Promoters can also act as one of the basic regulation mechanisms for the gene the precede. They can control how much the gene is expressed, and subsequently how much protein is produced. Certain sequences can accompany promoters and allow regulatory proteins to bind around promoters, further regulating the expression of the gene they preceded.
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        Synthetic Biology is still a new field with many opportunities for characterization and regulation. The discovery that DNA could be transferred between organisms occurred in 1946, and the first plant was genetically modified in 1983 (1). Since then, most synthetic biology has dealt with bacteria rather than plants. Bacteria are prokaryotic, simple organisms and the procedures for their genetic modification are shorter. Consequently, eukaryotic organisms, especially plants, have an enormous amount of untapped potential  (2).  Genetic “parts” for plant organisms are limited and often poorly characterized.  Under-developed parts prevent many great scientific ideas for plants from ever getting off the ground.  
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<img style="border-radius: 10%; margin: 20px"src="http://www.aei.org/files/2013/07/03/img-labplantshutterstock_085533189991.gif_item_large.jpg">
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Understanding how different promoters regulate can be useful for tailoring the expression of genes in a synthetic system. Promoter have been categorized before for bacteria. An example of this is the J23100 family of promoters in the parts registry which allow someone to control the expression of their genetic circuit. The goal of this project was to explore promoter expression in plants.
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Among the limitations of working with plants Penn State iGEM noticed that only one promoter was commonly available for expression of proteins in plants: CMV 35s promoter. Since in Eukaryotic organisms the promoter is currently the primary source of variable expression, a wider variety of characterized promoters would be very useful. Penn State iGEM aims to modestly characterize several plant promoters for enhanced expression regulation in plant synthetic biology.
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<h2 style="color: green" ID="Back"> Background</h2>
<h2 style="color: green" ID="Back"> Background</h2>
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Eukaryotic organisms tend to be larger more complex units than prokaryotes. Some of the differences include the presence of a nucleus, many cytoskeletal elements, and certain genetic devices (1). The diagram below shows the difference from a synthetic biology perspective (2).
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<img style="margin: 20px" src="http://www.nature.com/scitable/content/ne0000/ne0000/ne0000/ne0000/95221497/synbio_2_1.png">
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<img style="margin: 20px; margin-left: 100px" src="http://www.nature.com/scitable/content/ne0000/ne0000/ne0000/ne0000/95221522/synbio_2_1.png">
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Transcriptions factors, chromosome insertion, the 5- UTR, and the reporter gene itself all can play a role in the protein expression.  Although, the promoter region might be the most common and simplest way to modify protein expression in eukaryotes. 
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<img style="margin: 20px" src="http://images-mediawiki-sites.thefullwiki.org/08/1/4/3/76469642612625177.png">
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Although simple to use, Eukaryotic promoters are fairly complicated.  Their function is to call on a RNA polymerase to initiate transcription.  The ability of a promoter region to call in an RNA polymerase will determine its strength and the relative protein expression.  The are a number of factors involved in this process ,but it varies from promoter to promoter. 
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Although simple to use, Eukaryotic promoters are fairly complicated.  Their function is to call on a RNA polymerase to initiate transcription.  The ability of a promoter region to call in an RNA polymerase will determine its strength and the relative protein expression.  The are a number of factors involved in this process ,but it varies from promoter to promoter. 
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<h2 style="color: green" ID="Meth"> Method </h2>
<h2 style="color: green" ID="Meth"> Method </h2>

Revision as of 14:13, 6 August 2013

Plant Promoter Project

As plants are still novel organisms for most of synthetic biology, we we are interested in developing methods of control for our projects. Currently the Cauliflower Mosaic Virus 35S promoter is the most widely used plant promoter. In hopes of increasing the availability of plant promoters, our project aims at testing viral promoters due to their relative efficiency, as well as cytoskeletal protein promoters due to their natural abundance. Testing these promoters in parallel with the CaMV 35S will create a plant promoter catalog which can be used for future iGEMers exploration of plant synthetic biology.

Introduction

Synthetic Biology is still a new field with many opportunities for characterization and regulation. The discovery that DNA could be transferred between organisms occurred in 1946, and the first plant was genetically modified in 1983 (1). Since then, most synthetic biology has dealt with bacteria rather than plants. Bacteria are prokaryotic, simple organisms and the procedures for their genetic modification are shorter. Consequently, eukaryotic organisms, especially plants, have an enormous amount of untapped potential (2). Genetic “parts” for plant organisms are limited and often poorly characterized. Under-developed parts prevent many great scientific ideas for plants from ever getting off the ground.

Among the limitations of working with plants Penn State iGEM noticed that only one promoter was commonly available for expression of proteins in plants: CMV 35s promoter. Since in Eukaryotic organisms the promoter is currently the primary source of variable expression, a wider variety of characterized promoters would be very useful. Penn State iGEM aims to modestly characterize several plant promoters for enhanced expression regulation in plant synthetic biology.

Background

Eukaryotic organisms tend to be larger more complex units than prokaryotes. Some of the differences include the presence of a nucleus, many cytoskeletal elements, and certain genetic devices (1). The diagram below shows the difference from a synthetic biology perspective (2).

Transcriptions factors, chromosome insertion, the 5- UTR, and the reporter gene itself all can play a role in the protein expression. Although, the promoter region might be the most common and simplest way to modify protein expression in eukaryotes.

Although simple to use, Eukaryotic promoters are fairly complicated. Their function is to call on a RNA polymerase to initiate transcription. The ability of a promoter region to call in an RNA polymerase will determine its strength and the relative protein expression. The are a number of factors involved in this process ,but it varies from promoter to promoter.

Although simple to use, Eukaryotic promoters are fairly complicated. Their function is to call on a RNA polymerase to initiate transcription. The ability of a promoter region to call in an RNA polymerase will determine its strength and the relative protein expression. The are a number of factors involved in this process ,but it varies from promoter to promoter.

Method

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Results

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Discussion

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Further Study

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