Master Degree / Yüksek Lisans Tezleri

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

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Department: search.filters.department.Thesis (Master)--İzmir Institute of Technology, BioengineeringSubject: search.filters.subject.Escherichia coli

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  • Master Thesis
    Development of Gold Nanoparticle-Based Plasmonic Assay Platform for Esherichia Coli Detection
    (Izmir Institute of Technology, 2017) Erdoğan, Duygu; Yıldız, Ümit Hakan; Arslan Yıldız, Ahu; Yıldız, Ümit Hakan; Arslan Yıldız, Ahu; 04.01. Department of Chemistry; 03.01. Department of Bioengineering; 03. Faculty of Engineering; 04. Faculty of Science; 01. Izmir Institute of Technology
    The traditional methods for pathogen detection have long detection time and insufficient sensitivity. Optical methods can overcome these drawbacks. There are solution based nanoparticle growth in the literature to enhance a surface sensitivity for biosensing applications. In this project, surface refractive index (RI) sensitivity was enhanced on solid support via gold growth to develop a label free, simple and costeffective methodology for bacteria screening. The gold nanoparticles (GNPs) were grown on solid support by using 20 μl of HAuCl4 / 80 μl of NH2OH at varied incubation times. Firstly, about 20 nm GNPs were synthesized and immobilized on polystyrene surfaces. Then, these GNPs were utilized as seed particles, and grown on solid support. During GNPs growth, a red shift in the plasmonic wavelength was observed. Morphological characterization showed that almost uniform gold growth could be achieved. The plasmonic platform sensitivity was validated by varied concentrations of sucrose, ethanol and BSA solutions, showing that the plasmonic platform gave a response to any small RI change. Next, two different E.coli bacterial strains’ adsorption was tested. Adsorption screenings for about 107 E.coli DH5-alpha cells/ml and 107 E.coli BL21(DE3) cells/ml in Phosphate Buffer Saline were made on growth gold surfaces. Further, E.coli BL21(DE3) containing milk and apple juice were also adsorbed on these gold surfaces with a 30 min incubation time. The results showed that these gold surfaces exhibit higher binding kinetics for bacteria. Therefore, the proposed LSPR-based label free methodology can be an alternative to the bacteria screening in water or food samples.
  • Master Thesis
    Cloning and Expression of the Pseudomonas Ke38 Extra-Cellular Lipase Gene in E. Coli
    (Izmir Institute of Technology, 2013) Karakaş, Fulya; Arslanoğlu, Alper; Arslanoğlu, Alper; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of Technology
    Lipases are serine hydrolases that catalyze both the hydrolysis and synthesis of insoluble or poorly soluble long-chain triacylglycerols with an acyl chain length ≥ 10 carbon atoms based on the presence or absence of water. Lipases are produced and secreted by all kingdoms of life that are eukaryotes including plants, animals, fungi and prokaryotes including bacteria and archaea. However, microbial lipases, especially from bacteria, more useful than their plant and animal derivatives because of several important properties. Because of their acitivities in both aqueous and nonaqueous environments, lipases have specific applications in industry and medicine. The primary goals of this thesis were to clone and express the extra-cellular lipase gene from Pseudomonas sp. KE38, isolated from soil samples of Erciyes mountain in Kayseri, in E. coli and partial purification of the gene product. To achieve this aim, genome walking technique was used to obtain lipase gene from Pseudomonas sp. KE38, that gene was then cloned into pET28a expression vector and expressed in E. coli. The lipase expression of E. coli BL21 and its activity was screened with olive oil-Rhodamin B plate assay. After expression recombinant lipase was partially purified via inclusion body isolation. Moreover the optimum lipase production time of E. coli BL21 cells were determined and analyzed with SDS-PAGE. According to SDS-PAGE analysis the recombinant lipase was approximately 64 kDa and lipase production reached to the highest level after two hours of IPTG induction. As conclusion, recombinant lipase from Pseudomonas sp. KE38 was cloned into E. coli, expressed and partially purified.