Time has been flying at OIMB! This week has been particularly busy for me, although not with injections and imaging this time. I’ve been working on the molecular biology aspects of my project, specifically in terms of mutating Rock binding sites in Ect2, amplifying full length bat star Rock, and attempting to stitch together a purple urchin Rock. The purple urchin Rock has been as prickly and unyielding as the urchins themselves; after many failed PCR attempts, we decided to abandon hope of obtaining the full sequence in one go. Instead, we will combine the pieces that we have already obtained using a technique called Gibson assembly. Additionally, we will tag the N-terminal (front half) kinase domain with mCherry (a red fluorescent protein) and the C-terminal (tail half) regulatory domain with GFP. Both of these constructs may be valuable despite being separated from their other halves; naturally, the regulatory domain is constantly chasing the active kinase domain, so we have been referring to the kinase domain as the ‘cat’, and the regulatory domain as the ‘dog’. A few of this week's sunsets. Left: sunset at Bastendorff Beach, right: sunset at the OIMB beach. In addition to battling the purple urchin Rock, we have been working hard on mutating the Rock binding sites in Ect2. After using our gene blocks to mutate the Rock binding sites and amplify the new version of Ect2, we transformed the purified PCR product into E. coli and let the little bacteria make many copies of the plasmids we want. We then extracted the plasmids and conducted several digests with restriction enzymes that cut in known locations to confirm that the E. coli actually amplified what we wanted. Although the digests looked a little weird, we sent some of the purified plasmid DNA for sequencing to verify the presence of our constructs. Despite the setbacks I've described, life inside and outside of the lab has been fun. The REUs took a weekend trip up to Newport last weekend to visit the Oregon Coast Aquarium (see photos above). Watching the rays and skates swim above me in the "Passages of the Deep" tunnels was magical, as was watching the octopus feeding at the Hatfield Marine Science Center. I look forward to continuing exploring the Oregon coast and hopefully getting back some sequences this weekend to start injecting my probes next week!
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AuthorI am a rising junior at Carleton College, majoring in biology with minors in Neuroscience and Russian. I'm very excited to be working in Dr. George von Dassow's lab this summer, where I will be studying cell biology and embryology of marine invertebrates. Archives
August 2019
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