Team:Cornell/project/wetlab/fungal toolkit/agrobacterium
From 2013.igem.org
(Difference between revisions)
Line 17: | Line 17: | ||
<div class="twelve columns"> | <div class="twelve columns"> | ||
<h3>Background</h3> | <h3>Background</h3> | ||
- | 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 <i>Agrobacterium tumefaciens</i>, an organism that is able to transfer part of its DNA, known as T-DNA. Genetic constructs can be inserted into these T-DNA vectors and then transformed into <i>A. tumefaciens</i>, allowing it to insert this construct into a plant or fungal genome [1]. | + | 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 <i>Agrobacterium tumefaciens</i>, 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 <i>A. tumefaciens</i>, allowing it to insert this construct into a plant or fungal genome [1]. |
<br> | <br> | ||
<h3>Methods</h3> | <h3>Methods</h3> | ||
- | In order to transform our genetic constructs into <i>Ganoderma lucidum</i>, we first inserted them into pOSCAR, a T-DNA vector that contains regions that <i>A. tumefaciens</i> | + | In order to transform our genetic constructs into <i>Ganoderma lucidum</i>, we first inserted them into pOSCAR, a T-DNA vector that contains regions that <i>A. tumefaciens</i> can insert into fungal cells. Following transformation of <i>A. tumefaciens</i> with our constructs in the pOSCAR vector, we co-cultured the cells with <i>G. lucidum</i> in induction media that provides optimal conditions for T-DNA insertion for a period of 2 days. <i>G. lucidum</i> was then placed on a selective plate to isolate transformants [2]. |
</div> | </div> | ||
</div> | </div> |
Latest revision as of 21:52, 27 October 2013
Agrobacterium
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.