Team:University of Alberta/Plant Project

From 2008.igem.org

(Difference between revisions)
(→Time line)
 
(13 intermediate revisions not shown)
Line 1: Line 1:
 +
{|  align="center"
 +
<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 ==
== IGEM's First Binary Vector ==
[[image:Binary Vector.png]]
[[image:Binary Vector.png]]
Line 5: 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.
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>
[[Growing Plants]]<br>
[[Selection of Primary Transformants]]<br>
[[Selection of Primary Transformants]]<br>
[[Transformation Protocol]]
[[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.
 +
 
 +
'''First Seed Pod of the project'''<br>
 +
[[image:first seed pods.jpg|200px]]
-
'''Week 1'''(Before Biobricks Arrive)<br>
+
==Journal Articles of Interest==
-
PCR our the resistance gene and promoter from the Pcambia vector<br>
+
===Plant Info===
-
Use Site directed mutagenesis to remove NOTI sites from the vector<br>
+
<li>'''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
 +
<Br>
 +
[http://mbe.oxfordjournals.org/cgi/content/full/20/5/735 Link]
-
'''Week 2'''(biobricks have arrived)<br>
+
===Cell Free===
-
Digest new biobrick and confirm it is what it is.<br>
+
<li>'''Biosensor immunoassays for the detection of bisphenol A''': Gerardo R. Marchesinia, b, c, , , Eline Meulenberga, Willem Haasnootb and Hubertus Irthc
-
Insert Plant biobrick into the pCambia vector that has had the NOTI sites removed<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]
-
Transform Vector into Agrobacterium<br>
+
-
Confirm Transformation<br>
+
-
Create bulk culture of Agrobacterium for plant transformation
+
-
'''Week 3'''
+
<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>
-
Transform Plants ( That are 3 weeks old)<br>
+
-
Make another bulk culture for next weeks transformation <br>
+
-
'''Week 4-6'''
+
-
Keep transforming plants on tray every week to increase chances.<br>
+
-
Harvest seeds of transformed plants and replant them.(continue as seeds come into maturity)<br>
+
-
'''Week 6-End'''
+
-
select for homozygous plants
+

Latest revision as of 23:25, 29 October 2008

HOMEUAB.pngTEAMUA.pngPROJECTUA.pngPARTSUA.pngNOTEBOOKUA.png

Contents

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

Binary Vector.png

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
First seed pods.jpg

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
    Retrieved from "http://2008.igem.org/Team:University_of_Alberta/Plant_Project"