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- All Subjects: Biology
- Creators: School of Life Sciences
Differences between basic and applied research were explored through a wet-lab case study. Vaccinia virus (VACV) infections are a prime model of the competition between a virus and its host. VACV contains a gene that is highly evasive of the host immune system, gene E3L. The protein encoded by E3L is E3, which contains two highly conserved regions, a C-terminus, and a N-terminus. While the C-terminus is well-understood, the mechanism by which the N-terminus grants IFN resistance was previously unknown. This project demonstrated that the N-terminus prevents the initiation of programmed necrosis through host-encoded cellular proteins RIP3 and DAI. These findings provide insight into the function of the N-terminus of E3, as well as the unique functions of induced programmed necrosis.
This project was an example of “basic” research. However, it highlights the interconnectivity of basic and applied research and the danger in isolating both projects and perspectives. It points to the difficult decisions that must be made in science, and the need for a better research classification system that considers what makes science “good” outside of antiquated social class ideologies that have shaped science since ancient Greece. While there are no easy answers to determine what makes research “good,” thinking critically about the types of research projects that will be pursued, and the effects that research has on both science and society, will raise awareness, initiate new conversations, and encourage more dialogue about science in the 21st century.
The bull shark, Carcharhinus leucas, is a large species that it is commonly distributed worldwide in tropical and subtropical waters. Despite the bull sharks global distribution, little is known about its life history. In particular, the limited reproductive information suggests the bull shark is placental viviparous, assumed to have a biennial cycle, and that newborn pup nurseries are near the coast. In order to conserve and protect any species, an understanding of the habitats where reproductive events occur is needed. In order to better understand the habitat use in Biscayne bay, Fla, and whether certain areas are critical during the reproductive cycle of bull sharks, I will evaluate circulating levels of the hormones progesterone, estradiol, and testosterone using radioimmunoassay. These samples were collected by the University of Miami opportunistically between 2012-2020 shipped to Arizona State University, where they were analyzed. For my study a total of 73 mature samples, 27 females and 46 males, were collected over the sampling period. The results indicated that Biscayne bay is an important gestation area for bull sharks. The hormonal trends for males and females demonstrated an interesting reproductive cycle, which were further supported through other placental viviparous reproductive patterns. Females had a low level of estradiol throughout most of the year, besides in the summer where there were no sharks in the bay due to movement to estuaries. During their return to the bay, there was a peak in progesterone indicating early pregnancy. Male testosterone levels indicated that there was a production in sperm right before females speculated peak in estradiol.
Bermuda Land Snails make up a genus called Poecilozonites that is endemic to Bermuda and is extensively present in its fossil record. These snails were also integral to the creation of the theory of punctuated equilibrium. The DNA of mollusks is difficult to sequence because of a class of proteins called mucopolysaccharides that are present in high concentrations in mollusk tissue, and are not removed with standard DNA extraction methods. They inhibit Polymerase Chain Reactions (PCRs) and interfere with Next Generation Sequencing methods. This paper will discuss the DNA extraction methods that were designed to remove the inhibitory proteins that were tested on another gastropod species (Pomacea canaliculata). These were chosen because they are invasive and while they are not pulmonates, they are similar enough to Bermuda Land Snails to reliably test extraction methods. The methods that were tested included two commercially available kits: the Qiagen Blood and Tissue Kit and the Omega Biotek Mollusc Extraction Kit, and one Hexadecyltrimethylammonium Bromide (CTAB) Extraction method that was modified for use on mollusk tissue. The Blood and Tissue kit produced some DNA, the mollusk kit produced almost none, and the CTAB Extraction Method produced the highest concentrations on average, and may prove to be the most viable option for future extractions. PCRs attempted with the extracted DNA have all failed, though it is likely due to an issue with reagents. Further spectrographic analysis of the DNA from the test extractions has shown that they were successful at removing mucopolysaccharides. When the protocol is optimized, it will be used to extract DNA from the tissue from six individuals from each of the two extant species of Bermuda Land Snails. This DNA will be used in several experiments involving Next Generation Sequencing, with the goal of assembling a variety of genome data. These data will then be used to a construct reference genome for Bermuda Land Snails. The genomes generated by this project will be used in population genetic analyses between individuals of the same species, and between individuals of different species. These analyses will then be used to aid in conservation efforts for the species.