The Real Excitement Commences

Hello all!

Last time I blogged, I had successfully made my clones to work with! So as a review, I have my gene of interest, inserted into the plasmid that contains the binding domain and that construct is now in a specific yeast strain. I then looked through our lab’s database to see what genes people have cloned into the activation domain. After finding around 20 genes that others have cloned into the activation domain, I performed a separate transformation for each of the 20 genes, into the yeast strain containing my gene of interest, UBE3A (Also called the bait).  I then plated the transformants onto selective media and put the plates into a 30 degree incubator, where they will grow for 3-5 days. After the 3-5 day growth period is up, I will take a single colony from each plate and streak it onto another selective plate. If colonies grow after 3-5 days on the new selective media that indicates that a TRUE PROTEIN INTERACTION is occurring! I made a diagram that shows what I talked about, unfortunately, I could not get it to go directly on my blog post, but if you go into my gallery and click on “Pictorial Explanation of 3rd blog,” you can see the diagram! Here is the link- http://ccsummerresearch.blogs.wm.edu/files/2012/08/In-Yeast-Strain1.pdf