Team:Cornell/project/wetlab/fungal toolkit/fungal transformation

From 2013.igem.org

(Difference between revisions)
Line 22: Line 22:
                                         <h5>Methods</h5>
                                         <h5>Methods</h5>
                                         <h5>Transformants</h5>
                                         <h5>Transformants</h5>
 +
<div class="row">
 +
<div class="four columns">
 +
<center><img src = "https://static.igem.org/mediawiki/2013/0/09/Linear_plasmid_2.jpg"></center>
 +
text here
 +
</div>
 +
 +
<div class="four columns">
 +
<center><img src = "https://static.igem.org/mediawiki/2013/0/09/Linear_plasmid_2.jpg"></center>
 +
text here
 +
</div>
 +
                                <div class="four columns">
 +
<center><img src = "https://static.igem.org/mediawiki/2013/0/09/Linear_plasmid_2.jpg"></center>
 +
text here
 +
</div>
 +
</div>
<div class="row">
<div class="row">
<div class="six columns">
<div class="six columns">
-
<br><center><img src = "https://static.igem.org/mediawiki/2013/0/09/Linear_plasmid_2.jpg"></center>
+
<center><img src = "https://static.igem.org/mediawiki/2013/0/09/Linear_plasmid_2.jpg"></center>
</div>
</div>
<div class="six columns">
<div class="six columns">
-
<br><center><img src = ""></center>
+
<center><img src = "https://static.igem.org/mediawiki/2013/0/09/Linear_plasmid_2.jpg"></center>
</div>
</div>
</div>
</div>
-
Transformants of the pHG vector obtained from the Turgeon Lab. Conferred resistance against hygromycin.
+
text here
-
Transformants of the pNG vector obtained from the Turgeon Lab. Conferred resistance against geniticin.
+
-
<br><center><img src = ""></center>
+
-
Transformants of trpc promoter and GFP construct between <i>Cochlioblus heterostrophus</i> homology regions obtained from the pNG vector.
+
<h3>Agrobacterium Mediated Transformation</h3>
<h3>Agrobacterium Mediated Transformation</h3>
<h5>Background</h5>
<h5>Background</h5>

Revision as of 01:59, 29 October 2013

Cornell University Genetically Engineered Machines

Fungal Transformation

PEG Mediated Transformation

Background
Methods
Transformants
text here
text here
text here
text here

Agrobacterium Mediated Transformation

Background
Wide varieties of plant and fungal cells can be easily and efficiently transformed by agrobacterium-mediated transformation. In this process, plant or fungal tissue is co-cultured with Agrobacterium tumefaciens, an organism that is able to transfer part of its DNA to plants and fungi, known as T-DNA. Genetic constructs can be inserted into these T-DNA vectors and then transformed into A. tumefaciens, allowing it to insert this construct into a plant or fungal genome [1].
Methods
In order to transform our genetic constructs into Ganoderma lucidum, we first inserted them into pOSCAR, a T-DNA vector that contains regions that A. tumefaciens can insert into fungal cells. Following transformation of A. tumefaciens with our constructs in the pOSCAR vector, we co-cultured the cells with G. lucidum in induction media that provides optimal conditions for T-DNA insertion for a period of 2 days. G. lucidum was then placed on a selective plate to isolate transformants [2].

References

1. Michielse, Caroline B., Paul J. J. Hooykaas, Cees A. M. J. J. Hondel, and Arthur F. J. Ram. "Agrobacterium-mediated Transformation as a Tool for Functional Genomics in Fungi." Current Genetics 48.1 (2005): 1-17.

2. Paz, Zahi, María D. García-Pedrajas, David L. Andrews, Steven J. Klosterman, Lourdes Baeza-Montañez, and Scott E. Gold. "One Step Construction of Agrobacterium-Recombination-ready-plasmids (OSCAR), an Efficient and Robust Tool for ATMT Based Gene Deletion Construction in Fungi." Fungal Genetics and Biology (2011): 677-684.

Retrieved from "http://2013.igem.org/Team:Cornell/project/wetlab/fungal_toolkit/fungal_transformation"