Master Degree / Yüksek Lisans Tezleri
Permanent URI for this collectionhttps://hdl.handle.net/11147/3008
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Master Thesis Differentiation of Filamentous Fungi by Polymerase Chain Reaction (pcr) and Fourier Transform Infrared (ftir) Spectroscopy(Izmir Institute of Technology, 2017) Güngör, Sinem; Baysal, Ayşe Handan; Baysal, Ayşe Handan; 03.08. Department of Food Engineering; 03. Faculty of Engineering; 01. Izmir Institute of TechnologyFourier transform infrared (FTIR) spectroscopy is considered to be a rapid, reliable, sensitive, and a cost-effective technique, which could be used as an efficient tool for microorganism identification. Since bio-molecules, such as lipids, carbohydrates, and nucleic acids, have their own unique ‘vibrational’ fingerprints and characteristic functional groups, which correspond to specific infrared light frequencies, FTIR spectrum obtained for any compound gives the information on the unique ‘fingerprint’. The objective of this study was to investigate the ability of FTIR spectroscopy for differentiating different species of filamentous fungi. In this study, Erkence cultivar olives which were collected from different orchards were used for different fungal strain isolation. The fungi isolates were grown on Malt Extract Agar (MEA) and Czapek Yeast Agar (CYA) at room temperature of 25ºC for 10 days. 15 different genera and 53 species were identified by using Polymerase Chain Reaction (PCR) and characterized in terms of DNA sequencing. FTIR spectroscopy was applied to 71 species as a novel technique to identify fungi. 18 pre-defined species that were collected fom previous studies, were also used for FTIR spectroscopy investigation. Statistical analysis of the data was performed by using a principal component analysis (PCA). FTIR spectroscopy provides a potentially powerful approach to differentiate filamentous fungi.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.