Team:University of Alberta/Plant Project

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Latest revision as of 23:25, 29 October 2008

Introduction

To date, synthetic biology has focused largely on single cell organisms. The goal of our project is to build BioBrick to facilitate application of synthetic biology principles to the genetic engineering of higher plants. We are developing standardized, open source binary vectors, promoters, and resistance genes for use in plants, each of which is to conform to BioBrick specifications. The application of the BioBricks in transformation of protoplasts and whole plants of tobacco and Arabidopsis, respectively, will be demonstrated, as well as a general strategy for increasing the general availability of synthetic biology tools for plant genetic engineering.

IGEM's First Binary Vector

Our Binary Vector is a modified pCambia 0380 binary vector [link] We have added in our own poly linker which includes the proper prefix, suffix and primers to coincide with the IGEM regulations. We also added a CAM resistance cassette.

Useful Plant Protocols

Growing Plants
Selection of Primary Transformants
Transformation Protocol

Status of Project

Currently we have the polylinker and the vector that we are introducing it into (p0380). We have optimized the vector p0380 using site directed mutagenesis to remove all pre-existing notI sites from the vector. At the moment we are in the process of verifying that our polylinker has been successfully inserted into the vector. Due to the fact that we cannot say for sure if our polylinker has made it into the vector our vector will have to be submitted after the jamboree.

First Seed Pod of the project

Journal Articles of Interest

Plant Info

The Basic Helix–Loop–Helix Transcription Factor Family in Plants: A Genome-Wide Study of Protein Structure and Functional Diversity: Marc A. Heim, Marc Jakoby, Martin Werber, Cathie Martin, Bernd Weisshaar, and Paul C. Bailey
Link

Cell Free

Biosensor immunoassays for the detection of bisphenol A: Gerardo R. Marchesinia, b, c, , , Eline Meulenberga, Willem Haasnootb and Hubertus Irthc
Link

Basis of a High-Throughput Method for Nuclear Receptor Ligands: Tomohiko Kanayama, Satoru Mamiya, Tsutomu Nishihara and Jun-ichi Nishikawa J. Biochem. 133, 791–797 (2003)
Link

@@ Line 1: / Line 1: @@
-'''First Seed Pod of the project'''<br>
+{|  align="center"
-[[image:first seed pods.jpg|200px]]
+<span class="plainlinks">[{{SERVER}}{{localurl:Team:The_University_of_Alberta}} https://static.igem.org/mediawiki/2008/a/af/HOMEUAB.png]</span>
+<span class="plainlinks">[{{SERVER}}{{localurl:Team:The_University_of_Alberta/Team}} https://static.igem.org/mediawiki/2008/7/7c/TEAMUA.png]</span>
+<span class="plainlinks">[{{SERVER}}{{localurl:Team:The_University_of_Alberta/Project}} https://static.igem.org/mediawiki/2008/7/7e/PROJECTUA.png]</span>
+<span class="plainlinks">[{{SERVER}}{{localurl:Team:The_University_of_Alberta/Parts}} https://static.igem.org/mediawiki/2008/c/c0/PARTSUA.png]</span>
+<span class="plainlinks">[{{SERVER}}{{localurl:Team:The_University_of_Alberta/Notebook}} https://static.igem.org/mediawiki/2008/f/f9/NOTEBOOKUA.png]</span>
+{|  align="center"
+<br>
+==Introduction==
+To date, synthetic biology has focused largely on single cell organisms.  The goal of our project is to build BioBrick to facilitate application of synthetic biology principles to the genetic engineering of higher plants.  We are developing standardized, open source binary vectors, promoters, and resistance genes for use in plants, each of which is to conform to BioBrick specifications.  The application of the BioBricks in transformation of protoplasts and whole plants of tobacco and Arabidopsis, respectively, will be demonstrated, as well as a general strategy for increasing the general availability of synthetic biology tools for plant genetic engineering.
 == IGEM's First Binary Vector ==
 [[image:Binary Vector.png]]
@@ Line 8: / Line 16: @@
 We have added in our own poly linker which includes the proper prefix, suffix and primers to coincide with the IGEM regulations. We also added a CAM resistance cassette.
-==The First Plant Project Ever Submitted to IGEM==
+==Useful Plant Protocols==
-We hope to demonstrate that using biobricks in plants is not only useful but feasible in the restraints of this competition. We also hope to use the tools that we make to insert our toxin detection vector into plants.
-==Plant Protocol==
 [[Growing Plants]]<br>
 [[Selection of Primary Transformants]]<br>
 [[Transformation Protocol]]
-==Time line==
+==Status of Project==
-A new tray of plants will be planted every week.
+Currently we have the polylinker and the vector that we are introducing it into (p0380). We have optimized the vector p0380 using site directed mutagenesis to remove all pre-existing notI sites from the vector. At the moment we are in the process of verifying that our polylinker has been successfully inserted into the vector. Due to the fact that we cannot say for sure if our polylinker has made it into the vector our vector will have to be submitted after the jamboree.
-'''Week 1'''(Before Biobricks Arrive)<br>
+'''First Seed Pod of the project'''<br>
-PCR our the resistance gene and promoter from the Pcambia vector<br>
+[[image:first seed pods.jpg|200px]]
-Use Site directed mutagenesis to remove NOTI sites from the vector<br>
-'''Week 2'''(biobricks have arrived)<br>
+==Journal Articles of Interest==
-Digest new biobrick and confirm it is what it is.<br>
+===Plant Info===
-Insert Plant biobrick into the pCambia vector that has had the NOTI sites removed<br>
+<li>'''The Basic Helix–Loop–Helix Transcription Factor Family in Plants: A Genome-Wide Study of Protein Structure and Functional Diversity''':
-Transform Vector into Agrobacterium<br>
+Marc A. Heim, Marc Jakoby, Martin Werber, Cathie Martin, Bernd Weisshaar, and Paul C. Bailey
-Confirm Transformation<br>
+<Br>
-Create bulk culture of Agrobacterium for plant transformation
+[http://mbe.oxfordjournals.org/cgi/content/full/20/5/735 Link]
-'''Week 3'''
+===Cell Free===
-Transform Plants ( That are 3 weeks old)<br>
+<li>'''Biosensor immunoassays for the detection of bisphenol A''': Gerardo R. Marchesinia, b, c, , , Eline Meulenberga, Willem Haasnootb and Hubertus Irthc
-Make another bulk culture for next weeks transformation <br>
+<br>[http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TF4-4D04TNM-1&_user=1067472&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000051251&_version=1&_urlVersion=0&_userid=1067472&md5=4aab2bf3161d728c5c3bd69a3c418800 Link]
-'''Week 4-6'''
-Keep transforming plants on tray every week to increase chances.<br>
+<li>'''Basis of a High-Throughput Method for Nuclear Receptor Ligands''': Tomohiko Kanayama, Satoru Mamiya, Tsutomu Nishihara and Jun-ichi Nishikawa J. Biochem. 133, 791–797 (2003)<br>[http://jb.oxfordjournals.org/cgi/reprint/133/6/791?maxtoshow=&HITS=10&hits=10&RESULTFORMAT=1&author1=kanayama&author2=nishikawa&andorexacttitle=and&andorexacttitleabs=and&andorexactfulltext=and&searchid=1&FIRSTINDEX=0&sortspec=relevance&resourcetype=HWCIT Link]<br>
-Harvest seeds of transformed plants and replant them.(continue as seeds come into maturity)<br>
-'''Week 6-End'''
-select for homozygous plants
-<br>
-<br>
-<h2>'''Back to [https://2008.igem.org/Team:The_University_of_Alberta/Project the projects page].'''</h2>
-==Status of Project==
-Currently we have the multiple cloning site (MCS) and the vector that we are introducing it into (p0380). We have optimized the vector p0380 using site directed mutagenesis to remove all pre-existing notI sites from the vector. At the moment we are in the process of verifying that our MCS has been successfully inserted into the vector.