My first week at the UCSF lab was great and went extremely well!
Upon my arrival, on Monday, my mentor explained the 2 projects that I would be
working on over the course of the next few weeks. One project is based on an
inherited heart disease called ARVC and the other is based on the study of the
Calsequestrin protein. I have been working on both of these projects
simultaneously over the course of my first week here at the lab.
As part of the ARVC project, I’m still in the process of
creating a nuclease called Cel I which has the ability of basically cutting up
DNA at a specific target and ultimately detecting heterozygosity. I had to
extract CEL-I from celery juice and purify it using a protein purification protocol
my mentor gave me to follow. CEL-I, as I mentioned, is a nuclease the lab is
using to detect heterozygosity. Since ARVC is autosomal dominant, CEL-I helps
prove that an organism has successfully taken in the disease by running their
DNA onto a gel and seeing cut up pieces of DNA at a specific location.
The second project I have been working on is the
Calsequestrin project. I think of the CASQ2 project as climbing a ladder. Because
it deals with understanding the structure and function of a mutant protein, we
first begin with trying to understand the chemistry and physics of the mutant
protein, and this can be done in the test tube. Then we ask biology questions
about the protein, but in vitro. In vitro is learning about the protein in a
cell line that is not part of an animal. Afterwards we can look at what the
protein does in cell culture using mouse skeletal muscle cells that have been
modified to grow individually. Lastly we make an animal model and look for a
phenotype that shows evidence of the disease in vivo. I learned that this is
how any scientist would proceed for any project involving a mutant protein. You
start with the basic characterization up to the in vivo experiments. So far in this project, I have successfully
ligated our target DNA into a strain of bacteria and allowed a culture of
bacteria to grow overnight. The next day
I transformed the plasmid into competent cells called XL-1 cells and
miniprepped 5 different bacteria colonies and sent them for sequencing.
My lab bench
It’s only been a week here at the lab, and I’ve already
learned so much! I’m looking forward to the next couple weeks!
Azza J
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