Search results for: Yakup Uslu
19 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 49218 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 55517 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 81916 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 56015 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 96014 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 22413 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 14412 Hot Face of Cold War: 007 James Bond
Authors: Günevi Uslu Evren
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Propaganda is one of the most effective methods for changing individual and mass opinions. Propaganda tries to get the message across to people or masses to effect rather than to provide objective information. There are many types of propaganda. Especially, political propaganda is a very powerful method that is used by states during in both war and peace. The aim of this method is to create a reaction against them by showing within the framework of internal and external enemies. Propaganda can be practiced by many different methods. Especially during the Cold War Era, the US and USSR have tried to create an ideological effect by using the mass media intensively. Cinema, which is located at the beginning of these methods, is the most powerful weapon to influence the masses. In this study, the historical process of the Cold War is examined. Especially, these propagandas that had been used by United States and The Soviet Union were investigated. The purposes of propaganda and construction methods were presented. Cold War events and relations between the US and the USSR during the Cold War will be discussed. Outlooks of two countries to each other during the Cold War, propaganda techniques used defectively during Cold War and how to use the cinema as a propaganda tool will be examined. The film "From Russia with Love, James Bond 007" that was filmed in Cold War were examined to explain how cinema was used as a propaganda tool in this context.Keywords: cinema, cold war, James Bond, propaganda
Procedia PDF Downloads 51811 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 80210 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 10199 On the Network Packet Loss Tolerance of SVM Based Activity Recognition
Authors: Gamze Uslu, Sebnem Baydere, Alper K. Demir
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In this study, data loss tolerance of Support Vector Machines (SVM) based activity recognition model and multi activity classification performance when data are received over a lossy wireless sensor network is examined. Initially, the classification algorithm we use is evaluated in terms of resilience to random data loss with 3D acceleration sensor data for sitting, lying, walking and standing actions. The results show that the proposed classification method can recognize these activities successfully despite high data loss. Secondly, the effect of differentiated quality of service performance on activity recognition success is measured with activity data acquired from a multi hop wireless sensor network, which introduces high data loss. The effect of number of nodes on the reliability and multi activity classification success is demonstrated in simulation environment. To the best of our knowledge, the effect of data loss in a wireless sensor network on activity detection success rate of an SVM based classification algorithm has not been studied before.Keywords: activity recognition, support vector machines, acceleration sensor, wireless sensor networks, packet loss
Procedia PDF Downloads 4758 The Effect of Temperature and Salinity on the Growth and Carotenogenesis of Three Dunaliella Species (Dunaliella sp. Lake Isolate, D. salina CCAP 19/18, and D. bardawil LB 2538) Cultivated under Laboratory Conditions
Authors: Imen Hamed, Burcu Ak, Oya Işık, Leyla Uslu, Kubilay Kazım Vursavuş
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In this study, 3 species of Dunaliella (Dunaliella sp. Salt Lake isoalte (Tuz Gölü), Dunaliella salina CCAP19/18, and Dunaliella bardawil LB 2538) and their optical density, dry matter, chlorophyll a, total carotenoids, and β-carotene production were investigated in a batch system. The aim of this research was to compare carotenoids, and β-carotene production were investigated in a batch those 3 species. Therefore 2 stress factors were used: 2 different temperatures (20°C and 30°C) and 2 different salinities (30‰, and 60‰) were tested over a 17-day study. The highest growth and chlorophyll a was reported for Dunaliella sp. under 20°C/30‰ and 20°C/60‰ conditions respectively followed by D. bardawil and D. salina. Significant differences were noticed (p<0.05) for the other 3 species. The growth decreased as temperature and salinity increased since the lowest growth was noticed for the 30°C/60‰ group. The chlorophyll a content decreased also as temperature increased however when the NaCl concentration increased an augmentation of the content was noticed . In the 17th day of experiment the highest carotenoids concentration was reported for D. bardawil 20°C/30‰ (65,639±0,400 μg.mL−1) and the most important β carotene concentration was for D. salina 20°C/60‰ (8,98E-07±0,013 mol/L).Keywords: Dunaliella sp., Dunaliella salina, Dunaliella bardawil, growth, pigments, stress factors
Procedia PDF Downloads 3147 Mathematical Modelling of Ultrasound Pre-Treatment in Microwave Dried Strawberry (Fragaria L.) Slices
Authors: Hilal Uslu, Salih Eroglu, Betul Ozkan, Ozcan Bulantekin, Alper Kuscu
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In this study, the strawberry (Fragaria L.) fruits, which were pretreated with ultrasound (US), were worked on in the microwave by using 90W power. Then mathematical modelling was applied to dried fruits by using different experimental thin layer models. The sliced fruits were subjected to ultrasound treatment at a frequency of 40 kHz for 10, 20, and 30 minutes, in an ultrasonic water bath, with a ratio of 1:4 to fruit/water. They are then dried in the microwave (90W). The drying process continued until the product moisture was below 10%. By analyzing the moisture change of the products at a certain time, eight different thin-layer drying models, (Newton, page, modified page, Midilli, Henderson and Pabis, logarithmic, two-term, Wang and Singh) were tested for verification of experimental data. MATLAB R2015a statistical program was used for the modelling, and the best suitable model was determined with R²adj (coefficient of determination of compatibility), and root mean square error (RMSE) values. According to analysis, the drying model that best describes the drying behavior for both drying conditions was determined as the Midilli model by high R²adj and low RMSE values. Control, 10, 20, and 30 min US for groups R²adj and RMSE values was established as respectively; 0,9997- 0,005298; 0,9998- 0,004735; 0,9995- 0,007031; 0,9917-0,02773. In addition, effective diffusion coefficients were calculated for each group and were determined as 3,80x 10⁻⁸, 3,71 x 10⁻⁸, 3,26 x10⁻⁸ ve 3,5 x 10⁻⁸ m/s, respectively.Keywords: mathematical modelling, microwave drying, strawberry, ultrasound
Procedia PDF Downloads 1536 Honor Endorsement from the Perspective of System Justification and Regulatory Focus Orientation
Authors: Gülçin Akbas Uslu
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Honor cultures put importance on the sexual purity of women. Women are expected to avoid acts that may spoil their honor. The emphasis on honor leads to the subordination of women and the dominance of men. In order to protect and clean honor, women are exposed to physical and psychological violence. Therefore, understanding the motivations driving people to endorse honor bears importance. For this purpose, this study aims to explore honor endorsement from the joint perspective of System Justification Theory (SJT) and Regulatory Focus Theory (RFT). SJT asserts that people have a tendency to support and rationalize the system. The motivation to maintain the system may be a factor in the endorsement of honor. RFT proposes two distinct regulatory processes, namely promotion and prevention focus. Having a dominant prevention focus, such as a deep concern for responsibilities, risk avoidance, and minimizing negative outcomes, may have a role in honor. Data were collected conveniently from 366 participants (216 women; 150 men). Participants filled out Honor Endorsement Index, Honor Based System Justification Scale and Regulatory Focus Orientation Scale Results revealed that both regulatory focus and system justification play a role in understanding honor. One-way ANOVA findings showed that individuals with a dominant prevention focus endorse honor beliefs more than individuals with a dominant promotion focus. Besides, regression analysis revealed that prevention focus and system justification significantly and positively predict honor. Results provide clarifications for why honor has an important meaning in individuals’ life and why honor-based violence is approved. These findings bear great importance in Turkey, where emphasis on honor is high and can be used in reducing people’s adherence to honor, which is based on women’s sexuality and men’s power over women.Keywords: honor, system justification theory, regulatory focus theory, prevention focus
Procedia PDF Downloads 1535 Challenges in Achieving Profitability for MRO Companies in the Aviation Industry: An Analytical Approach
Authors: Nur Sahver Uslu, Ali̇ Hakan Büyüklü
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Maintenance, Repair, and Overhaul (MRO) costs are significant in the aviation industry. On the other hand, companies that provide MRO services to the aviation industry but are not dominant in the sector, need to determine the right strategies for sustainable profitability in a competitive environment. This study examined the operational real data of a small medium enterprise (SME) MRO company where analytical methods are not widely applied. The company's customers were divided into two categories: airline companies and non-airline companies, and the variables that best explained profitability were analyzed with Logistic Regression for each category and the results were compared. First, data reduction was applied to the transformed variables that went through the data cleaning and preparation stages, and the variables to be included in the model were decided. The misclassification rates for the logistic regression results concerning both customer categories are similar, indicating consistent model performance across different segments. Less profit margin is obtained from airline customers, which can be explained by the variables part description, time to quotation (TTQ), turnaround time (TAT), manager, part cost, and labour cost. The higher profit margin obtained from non-airline customers is explained only by the variables part description, part cost, and labour cost. Based on the two models, it can be stated that it is significantly more challenging for the MRO company, which is the subject of our study, to achieve profitability from Airline customers. While operational processes and organizational structure also affect the profit from airline customers, only the type of parts and costs determine the profit for non-airlines.Keywords: aircraft, aircraft components, aviation, data analytics, data science, gini index, maintenance, repair, and overhaul, MRO, logistic regression, profit, variable clustering, variable reduction
Procedia PDF Downloads 314 Profiling of Apoptotic Protein Expressions after Trabectedin Treatment in Human Prostate Cancer Cell Line PC-3 by Protein Array Technology
Authors: Harika Atmaca, Emir Bozkurt, Latife Merve Oktay, Selim Uzunoglu, Ruchan Uslu, Burçak Karaca
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Microarrays have been developed for highly parallel enzyme-linked immunosorbent assay (ELISA) applications. The most common protein arrays are produced by using multiple monoclonal antibodies, since they are robust molecules which can be easily handled and immobilized by standard procedures without loss of activity. Protein expression profiling with protein array technology allows simultaneous analysis of the protein expression pattern of a large number of proteins. Trabectedin, a tetrahydroisoquinoline alkaloid derived from a Caribbean tunicate, Ecteinascidia turbinata, has been shown to have antitumor effects. Here, we used a novel proteomic approach to explore the mechanism of action of trabectedin in prostate cancer cell line PC-3 by apoptosis antibody microarray. XTT cell proliferation kit and Cell Death Detection Elisa Plus Kit (Roche) was used for measuring cytotoxicity and apoptosis. Human Apoptosis Protein Array (R&D Systems) which consists of 35 apoptosis related proteins was used to assess the omic protein expression pattern. Trabectedin induced cytotoxicity and apoptosis in prostate cancer cells in a time and concentration-dependent manner. The expression levels of the death receptor pathway molecules, TRAIL-R1/DR4, TRAIL R2/DR5, TNF R1/TNFRSF1A, FADD were significantly increased by 4.0-, 21.0-, 4.20- and 11.5-fold by trabectedin treatment in PC-3 cells. Moreover, mitochondrial pathway related pro-apoptotic proteins Bax, Bad, Cytochrome c, and Cleaved Caspase-3 expressions were induced by 2.68-, 2.07-, 2.8-, and 4.5-fold and the expression levels of anti-apoptotic proteins Bcl-2 and Bcl-XL were reduced by 3.5- and 5.2-fold in PC-3 cells. Proteomic (antibody microarray) analysis suggests that the mechanism of action of trabectedin may be exerted via the induction of both intrinsic and extrinsic apoptotic pathways. The antibody microarray platform can be utilised to explore the molecular mechanism of action of novel anticancer agents.Keywords: trabectedin, prostate cancer, omic protein expression profile, apoptosis
Procedia PDF Downloads 4423 Determination of the Walkability Comfort for Urban Green Space Using Geographical Information System
Authors: Muge Unal, Cengiz Uslu, Mehmet Faruk Altunkasa
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Walkability relates to the ability of the places to connect people with varied destinations within a reasonable amount of time and effort, and to offer visual interest in journeys throughout the network. So, the good quality of the physical environment and arrangement of walkway and sidewalk appear to be more crucial in influencing the pedestrian route choice. Also, proximity, connectivity, and accessibility are significant factor for walkability in terms of an equal opportunity for using public spaces. As a result, there are two important points for walkability. Firstly, the place should have a well-planned street network for accessible and secondly facilitate the pedestrian need for comfort. In this respect, this study aims to examine the both physical and bioclimatic comfort levels of the current condition of pedestrian route with reference to design criteria of a street to access the urban green spaces. These aspects have been identified as the main indicators for walkable streets such as continuity, materials, slope, bioclimatic condition, walkway width, greenery, and surface. Additionally, the aim was to identify the factors that need to be considered in future guidelines and policies for planning and design in urban spaces especially streets. Adana city was chosen as a study area. Adana is a province of Turkey located in south-central Anatolia. This study workflow can be summarized in four stages: (1) environmental and physical data were collected by referred to literature and used in a weighted criteria method to determine the importance level of these data , (2) environmental characteristics of pedestrian routes gained from survey studies are evaluated to hierarchies these criteria of the collected information, (3) and then each pedestrian routes will have a score that provides comfortable access to the park, (4) finally, the comfortable routes to park will be mapped using GIS. It is hoped that this study will provide an insight into future development planning and design to create a friendly and more comfort street environment for the users.Keywords: comfort level, geographical information system (GIS), walkability, weighted criteria method
Procedia PDF Downloads 3092 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 3741 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 320