Internship Presentation

1. Internship Presentation Brendon Johnson 6/11/2010

2. Main Project: Transformation of Arabidopsis • The Main Project that I worked on over the Internship period was the transformation of Arabidopsis thaliana with Agrobacteriumtumefaciens, a procedure that is important to the research conducted in the lab.

3. How the Procedure Works • I first transformed competent cells with the DNA I wanted to eventually have expressed in the Arabidopsis. • Then engineered Agrobacterium accepts the T-DNA from the transformed E. Coli. • Arabidopsis’s floral tissue can be directly transformed with Agrobacterium, which makes it a better host plant to be transformed than some other plants

4. What Happens When Agrobacterium Transforms a Plant • Wild Type Agrobacteriumcontains a plasmid called pTI, standing for Tumor Inducing plasmid. • The difference between wild type Agrobacterium and engineered Agrobacterium used in the lab is the removal of the Oncogenicand Opine genes in the T-DNA region and the introduction of the genes that are being transformed into the plant.

5. Mechanism of Agrobacterium Gene Transfer Agrobacterium is able to transform plant cells because of two regions of DNA in the Tumor Inducing plasmid, the Viroperons, and the T-DNA region. The Viroperons code for the proteins that are essential to the transformation of the T-DNA region into the plant cells.