Master Degree / Yüksek Lisans Tezleri
Permanent URI for this collectionhttps://hdl.handle.net/11147/3008
Browse
2 results
Search Results
Master Thesis Investigation of Biomarkers Using Lipidome-Based Research Analysis in Sialidosis(01. Izmir Institute of Technology, 2024) Gümüş, İlker; Seyrantepe, Volkan; Seyrantepe, Volkan; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of TechnologyNeuraminidase 1, also known as N-acetyl-α-neuraminidase, is an enzyme found in lysosomes and encoded by the NEU1 gene. This enzyme is responsible for eliminating terminal sialic acids from glycoproteins and oligosaccharides. When mutations occur in the NEU1 gene, it leads to a particular lysosomal storage disorder known as sialidosis. Sialidosis is a rare genetic disorder that is inherited in an autosomal recessive manner. Sialidosis is classified into two subtypes: Type I, which has a later onset and Type II, which presents with early onset. In previous studies, an increase in glycolipid levels in visceral organs, and accumulation of sialyloligosaccharides and sialoglycoproteins were reported. However, the effect of NEU1 sialidase on secondary lipid expression levels in sialidosis pathology remains unknown. The relationship between lipid expression levels and inflammation of human and mice sialidosis fibroblasts cell lines was analyzed for the first time in this study. To understand the connection between secondary lipid alterations and inflammation in sialidosis molecular biological approaches and shotgun lipidome analysis were followed. The link between the secondary lipid alterations and its association with sialidosis was determined in our research. We have concluded that the findings not only provide the elucidation of the lipidome characteristics in sialidosis models of mice and patients, but they also have the potential to establish a connection between the lipidome features and the occurrence of neuroinflammation in sialidosis. Grasping the correlation between the changes in secondary lipids and inflammation may offer therapeutic strategies for sialidosis patients by modulating the expression of secondary lipids.Master Thesis Design, Construction and Expression of a Synthetic Gene for Metal Binding Proteins(Izmir Institute of Technology, 2009) Bal, Erhan; Şanlı Mohamed, Gülşah; Şanlı Mohamed, Gülşah; 04.01. Department of Chemistry; 04. Faculty of Science; 01. Izmir Institute of TechnologyConstruction of synthetic genes is today the most elegant way to optimize the heterologous expression of a recombinant protein. The availability of sequences of entire genome has significantly increased the number of protein targets which many of them will need to be overexpressed in cells other than the original source of DNA. Gene synthesis often provides a fast and economically efficient approach. In this study we have optimized a two-step polymerase chain reaction (2-step PCR) method for the fast and extremely accurate synthesis of a 186 bp CUP1 gene encoding yeast Saccharomyces cerevisiae copper metallothionein. A total of the six overlapping oligonucleotides ranged from 43 to 49 in length, designed with the unique restriction sites, were assembled in a single step PCR. The assembly was then further amplified by second PCR to produce a synthetic gene which has been cloned into the pET28a(+) vector to allow the expression of CUP1 gene in E. coli BL21 (DE3) host cell. In order to compare the difference in expression level of the gene with optimized codon usage for E. coli, CUP1gene was redesigned according to codon bias of host cell. A significant increase of expression level of codon optimized gene was obtained compared to original sequence of CUP1 gene of copper metallothionein in yeast Saccharomyces cerevisiae.