Matching Items (7)

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Cloning Sulfate Transporters in Growth Inhibited Yeast S. Cerevisiae

Description

The yeast project studies the growth of yeast Saccharomyces Cerevisiae (S. Cerevisiae) in high and low sulfate environments and analyzes the potential for genetically mutated plasmids to facilitate sulfate uptake

The yeast project studies the growth of yeast Saccharomyces Cerevisiae (S. Cerevisiae) in high and low sulfate environments and analyzes the potential for genetically mutated plasmids to facilitate sulfate uptake in gene deficient yeast medias. The goal of the project was to transform the Sul1 and Sul2 transporters into the nutrient deficient yeast strain BY4743 and observe growth in conditions that would otherwise prohibit growth in order to create a model that can be used to study the effect of sulfate concentration on the transporters. The experimental results showed that expressing the sulfate transporters in the BY4743 strain provided the potential for the yeast to grow in nutrient-poor media. The growth potential model allows for further analysis on the sulfate transporters and will be used for research projects going forward.

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Date Created
  • 2019-05

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The Development of a T Cell Receptor Expression System to Verify TCR Specificity of Expanded Clones from Whole Blood: The beginnings of Adoptive T cell Therapy and T Cell Receptor Prediction

Description

Immunology, the study of the immune system and its ability to distinguish self from non-self, is a rapidly advancing sector of molecular biology. Cancer, being host derived, provides a difficult

Immunology, the study of the immune system and its ability to distinguish self from non-self, is a rapidly advancing sector of molecular biology. Cancer, being host derived, provides a difficult challenge for immune cells to distinguish it from normal tissue. The historic treatment of cancer has had three main methods: radiation, chemotherapy, and surgery (1). Due to recent advancements in understanding the regulatory role of adaptive immunity against cancer, researchers have been attempting to engineer therapies to enhance patients’ immunities against their cancer. Immunotherapies, both passive and active, demonstrate potential for combating many diseases. Passive immunization provides temporary protection against a pathogen, whereas active immunization teaches the patient’s system to respond to the antigen independently, giving life-long immunity. Passive immunization, generally, is a much more expensive method of providing immunity and is commonly used in emergency situations. Anti-venom, for example, uses antibodies grown in lab to neutralize venom. Examples of active immunization are vaccines, which mimic the wild-type pathogen in a way that elicits an immune response, specifically naïve lymphocyte activation and maturation into memory lymphocytes. In terms of cancer therapy, both passive and active immunization are being tested for efficacy (2).

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Date Created
  • 2020-05

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Cloning Hepatitis B core-West Nile DIII DNA sequence into Gemini Viral Vector using Molecular Biology techniques.

Description

Virus-Like Particles (VLPs) are self-assembling structures that lack the viral genetic material. Therefore they are safer and more immunogenic than other forms of vaccines. The Hepatitis B core (HBc) VLPs

Virus-Like Particles (VLPs) are self-assembling structures that lack the viral genetic material. Therefore they are safer and more immunogenic than other forms of vaccines. The Hepatitis B core (HBc) VLPs are a novel mechanism through which delivery of DNA-based human vaccines are plausible. Production of VLPs require recombinant, rapidly replicating, plant-based systems such as the geminiviral replicon system. This project entails the cloning process of HBc-DIII fusion protein, a VLP that should form Domain III of the Envelope protein on West Nile Virus, into deconstructed geminiviral vector. The cloning process includes the HBc-DIII fusion protein DNA isolation, restriction enzyme digestion with NcoI and SacI, PCR changing the NcoI site on the HBc-DIII insert to XbaI, sequencing, ligation into geminiviral vector and transformation into an agrobacterium strain. The major impediment to the cloning process was the presence of multiple bands instead of the expected two bands while doing restriction enzyme digests. The troubleshooting process enabled speculating that due to the excess of restriction enzymes in the digestion volume, some of the DNA was not digested completely. Hence, multiple bands were observed. However, sequencing analysis and further cloning process ensured the presence of HBc-DIII insert band (approximately 800bp) in the Gemini vector. Lastly, the construct HBc-DIII in Gemini vector was ensured to be in agrobacterium for further experiments such as agro-infiltration.

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Date Created
  • 2016-05

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Glioblastoma in the Crosshairs: Development of a Dual Reporter Assay for Discovery of Olig2 Inhibiting Drugs

Description

A coincidence reporter construct, consisting of the p21-promoter and two luciferase genes (Firefly and Renilla), was constructed for the screening of drugs that might inhibit Olig2's tumorigenic role in glioblastoma.

A coincidence reporter construct, consisting of the p21-promoter and two luciferase genes (Firefly and Renilla), was constructed for the screening of drugs that might inhibit Olig2's tumorigenic role in glioblastoma. The reporter construct was tested using an Olig2 inhibitor, HSP990, as well as short hairpin RNA targeting Olig2. Further confirmatory analysis is needed before the reporter cell line is ready for high-throughput screening at the NIH and lead compound selection.

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Date Created
  • 2014-05

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Reproductive Cloning and the Perception of Individuality

Description

Reproductive cloning is the duplication of genetic material to reproduce a living organism. The sheep Dolly was the first adult mammal to be cloned and her birth unveiled a multitude

Reproductive cloning is the duplication of genetic material to reproduce a living organism. The sheep Dolly was the first adult mammal to be cloned and her birth unveiled a multitude of questions about the potential for cloning humans and how that might threaten human individuality. Given those questions, my project delves into how reproductive cloning relates to the idea of individuality across three subgroups: humans, utility animals such as those used for research or agriculture, and pets.

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Date Created
  • 2013-05

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The Hwang Woo-Suk Scandal and the Development of Bioethics in South Korea

Description

In 2004, the South Korean geneticist Woo-Suk Hwang published what was widely regarded as the most important research result in biotechnology of the year. In the prestigious American journal Science,

In 2004, the South Korean geneticist Woo-Suk Hwang published what was widely regarded as the most important research result in biotechnology of the year. In the prestigious American journal Science, he claimed that he had succeeded in cloning a human blastocyst, an embryo in its early stages (Hwang et al. 2004). A year later, in a second Science article, he made the earth-shattering announcement that he had derived eleven embryonic stem cell lines using his cloning technique (Hwang et al. 2005). The international scientific community was stunned. American scientists publicly fretted that President George W. Bush‘s 2001 executive order limiting federal funding for stem-cell research in the United States had put American bioscience behind the Koreans‘ (Paarlberg 2005). These breakthroughs offered potential solutions to immune system rejection of transplanted organs and possible cures for diseases such as rheumatoid arthritis, Parkinson‘s, Down‘s syndrome, and paralysis (Svenaeus 2007). However, within a year, Hwang was exposed as a fraud who had faked his results and pressured his female colleagues to donate eggs without informed consent. Despite protests against his methods from Korean religious and nongovernmental organizations, Hwang had used his prestige to ignore his ethical obligations. The Korean government, too, was slow to investigate Hwang and to subject his work to appropriate regulation.

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  • 2012-12

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An Investigation of the Formation, Function, and Future of Extrachromosomal Circular DNA

Description

Extrachromosomal circular DNA (eccDNA) has been identified in a broad range of eukaryotes and have been shown to carry genes and regulatory sequences. Additionally, they can amplify within a cell

Extrachromosomal circular DNA (eccDNA) has been identified in a broad range of eukaryotes and have been shown to carry genes and regulatory sequences. Additionally, they can amplify within a cell by autonomous replication or reintegration into the genome, effectively influencing copy number in cells. This has significant implications for cancer, where oncogenes are frequently amplified on eccDNA. However, little is known about the exact molecular mechanisms governing eccDNA functionality. To this end, we constructed a fluorescent reporter at an eccDNA-prone locus of the yeast genome, CUP1. It is our hope that this reporter will contribute to a better understanding of eccDNA formation and amplification within a cell.

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Date Created
  • 2021-05