Browsing by Author "Vignjevic, Mimi"
Now showing 1 - 2 of 2
Results Per Page
Sort Options
Item The development of the chick embryo heart and using it as a model for atrial septal research(2022) Vignjevic, Mimi; Davis, MonicaAs an essential model organism, chicks can be used to study embryonic development. Information obtained by experiments can be applied to human development to understand how humans develop, processes and mechanisms that occur during human development, and possible sources of developmental disorders. Due to the rapid development of chick hearts, and similar developmental mechanism to human hearts, experiments performed on chick hearts can be applied to human heart development and be used to study human developmental disorders. Atrial septal defect is a common heart defect present at birth in humans, causing a hole in the septum of the heart. Using chicks, researchers can identify how heart structures move to form the septal hole, what genetic mutations or teratogens produce the defect, and potential mechanisms and treatments that can be used to prevent or treat atrial septal defect. Ultimately, chick heart research provides a more in depth understanding of human heart development which further provides the scientific community a greater understanding of general embryonic development.Item Site-directed mutagenesis of position 204 threonine to isoleucine failed to generate acid-tolerant EGFP(2023) Vignjevic, Mimi; Harcombe, KimberleyProtonation of green fluorescent protein (GFP) in acidic conditions prevents the emission of fluorescent light and limits the ability to visualize, localize, and study acidic organelles. Therefore, it is critical to introduce mutations into enhanced GFP (EGFP) to generate acid- tolerant fluorescent proteins. This experiment aimed to replicate a threonine to isoleucine mutation at position 204 in EGFP and identify if acid-stable fluorescent proteins would be produced in the BL21(DE3) Escherichia coli system. Site-directed mutagenesis was utilized to generate T204I mutant EGFP. SDS-PAGE and fluorescence microscopy were employed to analyze induction success and fluorescence. Spectrofluorophotometry was used to determine the excitation and emission spectra of T204I mutant EGFP and whether acid-tolerant proteins were generated. Results illustrated that the mutation of threonine to isoleucine at position 204 produced fluorescent proteins at pH 7. However, at pH 6 and 5, proteins failed to fluoresce. The replication of the T204I mutation failed to generate acid-stable EGFP proteins in the BL21(DE3) E. coli system. There is significance in generating acid-stable cellular markers, as currently, no cellular markers thrive in acidic conditions. This limits the ability to study acidic organelles and acidic cellular processes. Creating acid-tolerant markers will permit a greater range of biological research.