The aim of study To purify GPCR from a local strain of S. cerevisiae using Ion exchange and gel filtration chromatography techniques , by packing materials for columns which will be chosen of low cost comparing to the already used in published researches, which depend on the costly affinity chromatography and other expensive methods of purification. Local strain of S. cerevisiae chosen for extraction and purification of G-protein coupled receptor (GPCR) .The strains were obtained from biology department in Al- Mosul University, Iraq. The isolated colony was activated on Yeast Extract Pepton Dextrose Broth (YEPDB) and incubated at 30 C˚ for 24 h .Loop fully of the yeast culture was transferred to (10ml) of yeast extract peptone glucose agar (YEPGA) slant , then incubated at 30C˚for 24h , after that it was stored at 4C˚ ,the yeast cultures were reactivated and persevered after each two weeks period. S.cerevisiae was identified by morphological, microscopic characterization and biochemical test . The GPCR that extract from whole cell of S.cerevisiae was purified by ion exchange chromatography using DEAE-Sepharose ,the bound proteins (negatively charged) were then eluted using gradient concentration of NaCl ranged between( 0.1 -0.5M). Gel filtration chromatography using Sepharose 6B was applied as a second step of purification. The optical density for each fraction was measured at 280 nm by UV-VS spectrophotometer then the GPCR concentration was determined by using ELISA Kit . The fractions which gave the highest absorbance and concentration of GPCR were collected .The molecular weight of GPCR was determined by gel filtration chromatography using blue dextrin solution. Standard curve was plotted between log of molecular weight for standard protein and the ratio of Ve/Vo of GPCR . The purity of the GPCR that extracted and purified from whole cell of S, cerevisiae were carried out by using SDS-PAGE electrophoresis . In ion exchange chromatography the fraction were collected with 5 ml tube at a flow rate 0.5 ml/ min and eluted with gradient (0.1-0.5M) of sodium chloride solution. Two proteins peaks appeared after eluted by the gradient concentration of sodium chloride, while no protein peaks appeared in the washing fractions. The GPCR concentration was measured in the fractions of these two protein peaks, data indicated that GPCR located in the first protein peak (eluted at 0.1M of NaCl) at fraction numbers between 3 and 9, the maximum concentration of GPCR was 9.281 with specific activity 71.58(ng/mg)protein , 3.125 purification folds and72.9(%) yield while the second peaks (eluted at 0.4 M of NaCl) don't give any concentration for GPCR, thus its neglected. Gel filtration chromatography was used as second step of purification which applied by using sepharose 6B. Results show single active protein peaks appeared that identical with the peak of GPCR at fractions numbers(29-35). The maximum concentration of GPCR was 9.082 (ng/ml)was observed in these fractions. The specific activity for these fractions was 151.37 (ng/mg) protein with 6.608 purification folds and 39.64 (%) yield. The present study a chive a relatively high purification of GPCR from whole cell of a local strain S. cerevisiae with fold purification 6.608 and a yield of 39.64 % and molecular weight about~33KD.
(1)
(1)
Investigating gender differences based on emotional changes becomes essential to understand various human behaviors in our daily life. Ten students from the University of Vienna have been recruited by recording the electroencephalogram (EEG) dataset while watching four short emotional video clips (anger, happiness, sadness, and neutral) of audiovisual stimuli. In this study, conventional filter and wavelet (WT) denoising techniques were applied as a preprocessing stage and Hurst exponent
(10)
(7)
Despite the global economic downturn, online e-commerce, especially the sale of counterfeit goods, continues to witness a rise in sales, and one of the important players in e-commerce is the owners of e-commerce auction sites. Although they do not have a direct role in trademark infringement, they still consider them indirectly responsible (secondary liability) for infringing on their trademarks. Thus, many cases have been filed by the owners of the original trademarks (which have been infringed by persons who have sold some of the goods and products that are imitating their marks through some electronic auctions) against the owners of those auctions. In view of the international character of the Internet, in different countries, and despit
... Show MoreCladophora and Spirulina algae biomass have been used for the removal of Tetracycline (TC) antibiotic from aqueous solution. Different operation conditions were varied in batch process, such as initial antibiotic concentration, different biomass dosage and type, contact time, agitation speed, and initial pH. The result showed that the maximum removal efficiencies by using 1.25 g/100 ml Cladophora and 0.5 g/100 ml Spirulina algae biomass were 95% and 94% respectively. At the optimum experimental condition of temperature 25°C, initial TC concentration 50 mg/l, contact time 2.5hr, agitation speed 200 rpm and pH 6.5. The characterization of Cladophora and Spirulina biomass by Fourier transform infrared (FTIR) indicates that the presenc
... Show More
(3)
(2)
(5)
(4)
The study aims to identify the concept of empowering women from the point of view of experts in the Palestinian society, specifically in Gaza, as well as to explore the foundations of their formation of this concept. Additionally, the study seeks to clarify the most important challenges facing the empowerment of women in Palestinian society. The study used the design of a grounded theory that seeks to build the theory through deep analysis of the data, as qualitative data were collected through holding two focus groups and six in-depth interviews with the study sample, who were selected by the method of targeted sampling. The sample included (16) individuals (9 female experts, 7 male experts) holding academic and community leadership pos
... Show MoreModified algae with nano copper oxide (CuO) were used as adsorption media to remove tetracycline (TEC) from aqueous solutions. Functional groups, morphology, structure, and percentages of surfactants before and after adsorption were characterised through Fourier-transform infrared (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy-dispersive spectroscopy (EDS). Several variables, including pH, connection time, dosage, initial concentrations, and temperature, were controlled to obtain the optimum condition. Thermodynamic studies, adsorption isotherm, and kinetics models were examined to describe and recognise the type of interactions involved. Resultantly, the best operation conditions were at pH 7, contact time
... Show More
(30)
(28)
Soil wetted pattern from a subsurface drip plays great importance in the design of subsurface drip irrigation (SDI) system for delivering the required water directly to the roots of the plant. An equation to estimate the dimensions of the wetted area in soil are taking into account water uptake by roots is simulated numerically using HYDRUS (2D/3D) software. In this paper, three soil textures namely loamy sand, sandy loam, and loam soil were used with three different types of crops tomato, pepper, and cucumber, respectively, and different values of drip discharge, drip depth, and initial soil moisture content were proposed. The soil wetting patterns were obtained at every thirty minutes for a total time of irrigation equ
... Show More
(9)
Modified algae with nano copper oxide (CuO) were used as adsorption media to remove tetracycline (TEC) from aqueous solutions. Functional groups, morphology, structure, and percentages of surfactants before and after adsorption were characterised through Fourier-transform infrared (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy-dispersive spectroscopy (EDS). Several variables, including pH, connection time, dosage, initial concentrations, and temperature, were controlled to obtain the optimum condition. Thermodynamic studies, adsorption isotherm, and kinetics models were examined to describe and recognise the type of interactions involved. Resultantly, the best operation conditions were at pH 7, contact time
... Show More
(28)