Master Degree / Yüksek Lisans Tezleri

Permanent URI for this collectionhttps://hdl.handle.net/11147/3008

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Subject: search.filters.subject.Cloning

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  • Master Thesis
    Cloning and Purification of Biosynthetic Enzymes
    (Izmir Institute of Technology, 2016) Gürelme, Ebru; Şanlı Mohamed, Gülşah; Sürmeli, Nur Başak; Sürmeli, Nur Başak; Şanlı Mohamed, Gülşah; 03.01. Department of Bioengineering; 04.01. Department of Chemistry; 03. Faculty of Engineering; 04. Faculty of Science; 01. Izmir Institute of Technology
    Bioproducts have become prominent with their sustainable, eco-friendly and renewable features. In recent years, research and development studies focused on production of biodiesel and pharmaceuticals. Biodiesel can be synthesized in the form of fatty acid ethyl ester (FAEE) by in vivo activity. This synthesis is catalyzed by wax ester synthases (WS). This study aims cloning and purification of wax ester synthases from Psychrobacter arcticus 273-4 (PaWES) and Mus musculus C57BL/6 (MmWES). PaWES was cloned and expressed by Escherichia coli BL21(DE3) strain, at the proper conditions with using pET expression system. It was purified with approximately 1 mg yield. Cloning of the second wax ester synthase (MmWES) was achieved to Saccha-romyces cerevisiae and it was purified with lower than 1 mg yield. The other aim of this study is related to taxadien-5α-ol-O-acetyltransferase from Taxus cuspidata (TcT5AT). This enzyme belongs to the biosynthesis pathway of Taxol, which is the most commonly used chemotherapy drug. Cloning and purification studies of this enzyme were successfully performed. It was expressed by Escherichia coli BL21(DE3) Star strain and purified with the yield of 23 mg. Immobilized Metal Affinity Chromatog-raphy (IMAC) is used for all three enzymes as a purification strategy. This project can pave the way for structural studies of all biosynthetic enzymes mentioned above. In summary, the findings of this study will circuitously help for solving the relationship between function and structure of these enzymes. It may lead to increased generation of Taxol and FAEE based biodiesel.
  • Master Thesis
    Cloning of Polysome-Associated Small Rnas in Drosophila Melanogaster Embryos
    (Izmir Institute of Technology, 2009) Yiğit, Hatice; Akgül, Bünyamin; Akgül, Bünyamin; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of Technology
    Genome-encoded regulatory small RNAs are classified into 3 groups; microRNAs (miRNAs), endogeneous small interfering RNAs (endo siRNAs) and piwi interacting RNAs (piRNAs). miRNAs, 17-21 nucleotide in size, are involved in posttranscriptional gene regulation via precise or imprecise base pairing with target mRNAs resulting in either target mRNA degradation or translational inhibition. Endo siRNAs ,on the other hand, may function transposon regulation but their precise regulatory function and mechanism have not been elucidated yet. piRNAs are mainly involved in transposon silencing in spermatogenesis. Despite their discovery, biological roles and modes of functions of small RNAs remain to be elucidated. The aim of this thesis was to identify polysome-associated small RNAs in Drosophila melanogaster embryos by deep sequencing and investigate their role in translational regulation. Deep sequencing and microarray results determined stage and fraction specific distribution of genome encoded small RNAs. Surprisingly, the results implied that mRNAs may be posttranscriptionally regulated by antisense transcripts in polysome.