Training Complete

For the past few weeks, Dr. Hinton has trained me and my fellow summer lab members on various procedures and experimental techniques that are commonly used in lab. We have learned how to properly use the tissue culture room, which is a sterile room where we are able to culture various cell lines. There, we are able to treat our cells with different plasmids, giving them different expressions, and under treatment, we can see how these different expressions react under treatments. Along with that we have learned how to make more plasmids when we are running low. This process can be extensive, especially when they are multiple plasmids that need to be replenished. We used E. Coli bacteria to help culture the plasmids. Once the bacteria was transformed with corresponding plasmids, we broke up the cytoskeleton and other extracellular membrane proteins  to isolate the plasmids for further experiments. We have also learned how to do Western blots. Western blots help detect certain proteins with the help of antibodies that are specific to the proteins of interest. Western blots are done by first running a 2D gel with an electric current. After that, the gel is placed into a machine to transfer the data onto a membrane which allows for ease with antibody tagging. Training has gone very well so far, and I cannot wait to start my independent project.

June 7-14 Pyruvate Kinase Inhibition

Use of Molecules to Inhibit Pyruvate Kinase

With a method to measure the activity of pyruvate kinase, the research could begin. Next, the goal was to investigate the effects of various molecules on the activity of pyruvate kinase. This had to be done in three stages. In the first stage, the molecule modifies pyruvate kinase. In the second stage pyruvate kinase is allowed to react. Finally, in the third stage, a simple reaction occurs to make the product from stage 2 measurable.

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Exploring the Roles of Pseudophosphatase MK-STYX in Autophagy

Exploring the Roles of Pseudophosphatase MK-STYX in Autophagy

 

Cells carry out an incredible variety of processes to keep themselves and the greater organism they comprise functioning efficiently and healthily. As with most complex systems, cells possess a means of eliminating unwanted components and reusing or recycling the individual parts of these superfluous components into important constituent parts like amino acids and nucleic acids. This cellular recycling system is known as autophagy, and it permits organisms and their various cell types to remove redundant biomolecules [1]. In addition, autophagy allows cells to sustain themselves in times of starvation or stress by breaking down complex organelles and proteins into energy and basic biological building blocks. Unsurprisingly, autophagy is highly relevant to both healthy and diseased states, because a baseline level of autophagy is essential to an organism’s physical wellbeing. Conversely, a grossly dysregulated autophagic process affects and mediates pathogenesis and progression of cancers, neurodegenerative diseases, and metabolic disorders [2]. The pseudophosphatase MK-STYX and its roles in cellular processes represent the primary research focus of the Hinton Lab.  As a pseudophosphatase, MK-STYX lacks the catalytic ability to remove a phosphate group from proteins, but can bind targets with its pseudophosphatase domain and protein-interacting CH2 domain.  The importance of pseudophospahtases has only recently been accepted, thus MK-STYX and the other proteins in this group populate an exciting forefront of scientific inquiry.  Previous findings from our research lab have demonstrated that MK-STYX activity helps clear stress granules (made of translationally halted mRNA and associated proteins) from cells [3].  The two pathways that clear stress granules are the autophagy and ubiquitin protease pathways.  Based on recent studies conducted by our lab that showed MK-STYX altering the activity and expression levels of autophagy proteins, I decided to focus my research efforts on further characterizing MK-STYX’s roles in this critical intracellular homeostatic process.

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Abstract: Social phenotypes in S. cerevisiae

Microbes were once thought to just float around at random not interacting with each other in a meaningful way, however we now know that they exhibit complex social interactions, such as cooperation, communalism, competition, and even chemical warfare. Saccharomyces cerevisiae is an extremely powerful model organism for studies of eukaryotic biology. It has been studied intensely, in great detail, for many years and has been used to unravel many of the basic processes underlying genetics and cell biology. Until the past 5 years, however, the study of its social interactions with other yeast has not been looked at in detail. My proposal aims to characterize an understudied social phenotype expressed by many wild S. cerevisiae strains in order to understand the functioning of cooperative yeast communities.

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Locative Media and QR Codes- Abstract

Locative media is a term first coined by Karlis Kalnins and is derived from a Latvian verb tense that declares a “final location of action” (Zeffiro, 2012). Christian Licoppe defines locative media as “any kind of networked service available on mobile terminals which is able to provide to users on the move some reflexive awareness of the location of themselves and or others, or of their mutual proximity” (Zeffiro, 2012).

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