Search results for: Yakup Ulusu
11 Modification of Escherichia coli PtolT Expression Vector via Site-Directed Mutagenesis
Authors: Yakup Ulusu, Numan Eczacıoğlu, İsa Gökçe, Helen Waller, Jeremy H. Lakey
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Besides having the appropriate amino acid sequence to perform the function of proteins, it is important to have correct conformation after this sequence to process. To consist of this conformation depends on the amino acid sequence at the primary structure, hydrophobic interaction, chaperones and enzymes in charge of folding etc. Misfolded proteins are not functional and tend to be aggregated. Cysteine originating disulfide cross-links make stable this conformation of functional proteins. When two of the cysteine amino acids come side by side, disulfide bond is established that forms a cystine bridge. Due to this feature cysteine plays an important role on the formation of three-dimensional structure of many proteins. There are two cysteine amino acids (C44, C69) in the Tol-A-III protein. Unlike protein disulfide bonds from within his own, any non-specific cystine bridge causes a change in the three dimensional structure of the protein. Proteins can be expressed in various host cells as directly or fusion (chimeric). As a result of overproduction of the recombinant proteins, aggregation of insoluble proteins in the host cell can occur by forming a crystal structure called inclusion body. In general fusion proteins are produced for provide affinity tags to make proteins more soluble and production of some toxic proteins via fusion protein expression system like pTolT. Proteins can be modified by using a site-directed mutagenesis. By this way, creation of non-specific disulfide crosslinks can be prevented at fusion protein expression system via the present cysteine replaced by another amino acid such as serine, glycine or etc. To do this, we need; a DNA molecule that contains the gene that encodes for the target protein, required primers for mutation to be designed according to site directed mutagenesis reaction. This study was aimed to be replaced cysteine encoding codon TGT with serine encoding codon AGT. For this sense and reverse primers designed (given below) and used site-directed mutagenesis reaction. Several new copy of the template plasmid DNA has been formed with above mentioned mutagenic primers via polymerase chain reaction (PCR). PCR product consists of both the master template DNA (wild type) and the new DNA sequences containing mutations. Dpn-l endonuclease restriction enzyme which is specific for methylated DNA and cuts them to the elimination of the master template DNA. E. coli cells obtained after transformation were incubated LB medium with antibiotic. After purification of plasmid DNA from E. coli, the presence of the mutation was determined by DNA sequence analysis. Developed this new plasmid is called PtolT-δ.Keywords: site directed mutagenesis, Escherichia coli, pTolT, protein expression
Procedia PDF Downloads 37410 Fabrication of Electrospun Green Fluorescent Protein Nano-Fibers for Biomedical Applications
Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz
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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.Keywords: biomaterial, GFP, nano-fibers, protein expression
Procedia PDF Downloads 3209 Investigation of Heating Behaviour of E-Textile Structures
Authors: Hande Sezgin, Senem Kursun Bahadır, Yakup Erhan Boke, Fatma Kalaoğlu
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Electronic textiles (e-textiles) are fabrics that contain electronics and interconnections with them. In this study, two types of base yarns (cotton and acrylic) and three conductive steel yarns with different linear resistance values (14Ω/m, 30Ω/m, 70Ω/m) were used to investigate the effect of base yarn type and linear resistance of conductive yarns on thermal behavior of e-textile structures. Thermal behavior of samples were examined by thermal camera.Keywords: conductive yarn, e-textiles, smart textiles, thermal analysis
Procedia PDF Downloads 5558 A ZVT-ZCT-PWM DC-DC Boost Converter with Direct Power Transfer
Authors: Naim Suleyman Ting, Yakup Sahin, Ismail Aksoy
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This paper presents a zero voltage transition-zero current transition (ZVT-ZCT)-PWM DC-DC boost converter with direct power transfer. In this converter, the main switch turns on with ZVT and turns off with ZCT. The auxiliary switch turns on and off with zero current switching (ZCS). The main diode turns on with ZVS and turns off with ZCS. Besides, the additional current or voltage stress does not occur on the main device. The converter has features as simple structure, fast dynamic response and easy control. Also, the proposed converter has direct power transfer feature as well as excellent soft switching techniques. In this study, the operating principle of the converter is presented and its operation is verified for 1 kW and 100 kHz model.Keywords: direct power transfer, boost converter, zero-voltage transition, zero-current transition
Procedia PDF Downloads 8197 Assessing the Usability and Accessibility of Turkey E-Government Websites
Authors: Yakup Akgül, Kemal Vatansever
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E-Government services offer citizens an easy way to accomplish their work anytime and anywhere. Ensuring the accessibility and usability of such services is crucial to citizens to allow smooth online transaction. In this paper, an empirical study to investigate the accessibility and usability of a representative sample of Turkish e-government services presented. The study evaluated sixty one Turkish government websites according to four perspectives.The accessibility will be based on the compliance to WCAG 2.0 recommendations, and the usability will be based on a heuristics-based content, HTML and CSS validity and current use of HTML 5 and ARIA. The evaluation results show that the examined e-Government services have one or more accessibility issues. On the other hand, in terms of usability, most services were usable and well designed, thus little work is recommended to make them more user-friendly and attractive to citizen.Keywords: accessibility, e-government, usability, Turkey
Procedia PDF Downloads 5606 The Effects of Turkish Soap Operas on the Image of Turkey in the Middle Europe
Authors: Yakup Uslu
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The purpose of this study is to reveal how the Turkish soap operas effect the image of Turkey in the Middle Europe. In last decades, Turkish soap operas have been shown on TV in the middle European countries. A research based on face to face questioning was done in February and June 2014 in Slovakia and the Czech Republic. The participants were seven women and six men from the Czech Republic, 8 women and 6 men from Slovakia. According to results of the research, the Turkish image has been changed substantially after broadcasting the soap operas. In general, the Turkish soap operas have had positive effects on the image of Turkey. The other result of the study shows that most of the people in Slovakia and Czech Republic want to come to Turkey as tourists and want to visit the places where the soap operas have been shooted.Keywords: Turkish soap operas, image of Turkey, Slovakia, Czech Republic
Procedia PDF Downloads 4925 1 kW Power Factor Correction Soft Switching Boost Converter with an Active Snubber Cell
Authors: Yakup Sahin, Naim Suleyman Ting, Ismail Aksoy
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A 1 kW power factor correction boost converter with an active snubber cell is presented in this paper. In the converter, the main switch turns on under zero voltage transition (ZVT) and turns off under zero current transition (ZCT) without any additional voltage or current stress. The auxiliary switch turns on and off under zero current switching (ZCS). Besides, the main diode turns on under ZVS and turns off under ZCS. The output current and voltage are controlled by the PFC converter in wide line and load range. The simulation results of converter are obtained for 1 kW and 100 kHz. One of the most important feature of the given converter is that it has direct power transfer as well as excellent soft switching techniques. Also, the converter has 0.99 power factor with the sinusoidal input current shape.Keywords: power factor correction, direct power transfer, zero-voltage transition, zero-current transition, soft switching
Procedia PDF Downloads 9604 The Pomade for Treatment of Bovine Papilomavirus-Induced Warts in Teats
Authors: Mehmet Kale, Ramazan Sencan, Sibel Yavru, Ahmet Ak, Nuri Mamak, Sibel Hasırcıoglu, Mesih Kocamuftuoglu, Yakup Yıldırım, Hasbi Sait Saltık
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Bovine Papilloma Virus (BPV)-induced warts can cause mastitis, teat blindness, reduction of milk yield, udder deformities, and a difficulty in getting the teats into the milking machine. Especially, surgical operations cannot be performed in BPV-induced teat warts because of the increased sensitivity of the breast region and small-sized papillomas. Thus, there is a need to find new topical treatment methods. We have developed a pomade for treatment of BPV in cattle. The pomade is consists of lanoline, snakeskin (two special kind of snake), alcohol, vaseline, and ether. Firstly, we determined 46 cattle with teat warts. In the study, BPV antigen was detected in 28 cattle blood samples (61%) by ELISA. The pomade was applied to all BPV infected animals. The regression and recovery of warts were 100% in all animals. We advised using the pomade for treatment of BPV-induced warts in teats.Keywords: bovine papilloma virus, pomade, teat, udder
Procedia PDF Downloads 2243 A Patent Trend Analysis for Hydrogen Based Ironmaking: Identifying the Technology’s Development Phase
Authors: Ebru Kaymaz, Aslı İlbay Hamamcı, Yakup Enes Garip, Samet Ay
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The use of hydrogen as a fuel is important for decreasing carbon emissions. For the steel industry, reducing carbon emissions is one of the most important agendas of recent times globally. Because of the Paris Agreement requirements, European steel industry studies on green steel production. Although many literature reviews have analyzed this topic from technological and hydrogen based ironmaking, there are very few studies focused on patents of decarbonize parts of the steel industry. Hence, this study focus on technological progress of hydrogen based ironmaking and on understanding the main trends through patent data. All available patent data were collected from Questel Orbit. The trend analysis of more than 900 patent documents has been carried out by using Questel Orbit Intellixir to analyze a large number of data for scientific intelligence.Keywords: hydrogen based ironmaking, DRI, direct reduction, carbon emission, steelmaking, patent analysis
Procedia PDF Downloads 1442 A Single Phase ZVT-ZCT Power Factor Correction Boost Converter
Authors: Yakup Sahin, Naim Suleyman Ting, Ismail Aksoy
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In this paper, a single phase soft switched Zero Voltage Transition and Zero Current Transition (ZVT-ZCT) Power Factor Correction (PFC) boost converter is proposed. In the proposed PFC converter, the main switch turns on with ZVT and turns off with ZCT without any additional voltage or current stresses. Auxiliary switch turns on and off with zero current switching (ZCS). Also, the main diode turns on with zero voltage switching (ZVS) and turns off with ZCS. The proposed converter has features like low cost, simple control and structure. The output current and voltage are controlled by the proposed PFC converter in wide line and load range. The theoretical analysis of converter is clarified and the operating steps are given in detail. The simulation results of converter are obtained for 500 W and 100 kHz. It is observed that the semiconductor devices operate with soft switching (SS) perfectly. So, the switching power losses are minimum. Also, the proposed converter has 0.99 power factor with sinusoidal current shape.Keywords: power factor correction, zero-voltage transition, zero-current transition, soft switching
Procedia PDF Downloads 8021 A Soft Switching PWM DC-DC Boost Converter with Increased Efficiency by Using ZVT-ZCT Techniques
Authors: Yakup Sahin, Naim Suleyman Ting, Ismail Aksoy
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In this paper, an improved active snubber cell is proposed on account of soft switching (SS) family of pulse width modulation (PWM) DC-DC converters. The improved snubber cell provides zero-voltage transition (ZVT) turn on and zero-current transition (ZCT) turn off for main switch. The snubber cell decreases EMI noise and operates with SS in a wide range of line and load voltages. Besides, all of the semiconductor devices in the converter operate with SS. There is no additional voltage and current stress on the main devices. Additionally, extra voltage stress does not occur on the auxiliary switch and its current stress is acceptable value. The improved converter has a low cost and simple structure. The theoretical analysis of converter is clarified and the operating states are given in detail. The experimental results of converter are obtained by prototype of 500 W and 100 kHz. It is observed that the experimental results and theoretical analysis of converter are suitable with each other perfectly.Keywords: active snubber cells, DC-DC converters, zero-voltage transition, zero-current transition
Procedia PDF Downloads 1019