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.
The purpose of the research is to identify the role of Islamic banks in facing the financial crisis in Iraq for the period 2012-2014, and highlight the various factors that contributed to the emergence of the financial crisis, as well as to identify the problems and challenges facing the Islamic banking business in Iraq and identify the reasons and find appropriate solutions, And the development of treatments to avoid the occurrence of such problems and overcome them. The main findings of the research are as follows,Islamic banks (the sample of the study) were affected by the global financial crisis indirectly, as the financial crisis turned into an economic crisis affecting the real economy, because the activity of Islamic banks is more
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In this study, a packed bed was used to remove pathogenic bacteria from synthetic contaminated water. Two types of packing material substrates, sand and zeolite, were used. These substrates were coated with silver nanoparticles (AgNPs), which were prepared by decomposition of Ag ions from AgNO3 solution. The prepared coated packings were characterized using scanning electron microscopy, energy-dispersive X-ray spectroscopy and transmission electron microscopy. The packed column consisted of a PVC cylinder of 2 cm diameter and 20 cm in length. The column was packed with silver nanoparticlecoated substrates (sand or zeolite) at a depth of 10 cm. Four types of bacteria were studied: Escherichia coli, Shigella dysenteriae, Pseudomonas aerugi
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Mammary tumors (CMT) in dogs in Iraq may be induced by carcinogenic war ordnance. In our study, 10 virgin un- spayed military/pet bitches aged 5-15 years presented with abnormal masses in the abdomen with painful oedema, swelling, anorexia, weight loss, weakness and mild fever. Examination of regional lymph nodes and thoracic radiography confirmed metastasis. Tumors were excised and determined to be mostly adenocarcinomas involving multiple glands, solid in texture, 5-15 cm in size, mostly in the inguinal mammary glands at stage T3: >5 cm. Microscopy confirmed presence of adenocarcinoma in 8 dogs and solid carcinoma in 2 with half of tumors being grade III. Tumors had pleomorphic hyperchromatic cell nuclei in stroma, epithelial cells of duc
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Trickle irrigation is a system for supplying filtered water and fertilizer directly into the soil and water and it is allowed to dissipate under low pressure in an exact predetermined pattern. An equation to estimate the wetted area of unsaturated soil with water uptake by roots is simulated numerically using the HYDRUS-2D/3D software. In this paper, two soil types, which were different in saturated hydraulic conductivity were used with two types of crops tomato and corn, different values of emitter discharge and initial volumetric soil moisture content were assumed. It was assumed that the water uptake by roots was presented as a continuous sink function and it was introduced into Richard's equation in the unsaturated z
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knowledge management and managers have a primary school principals from the standpoint of their assistants M . Dr. Suad Vegetables Abbas Research problem: The administration school require a leader administratively The efficiency and skills of management and knowledge to make it effective in the administration follows the scientific method and knowledge of those who through administrative methods emanating from the trends of modern management in the management of educational , as well as the presence of the leaders of effective management has the knowledge and know-how administrative work and how to deal with others and investment optimized for all of the resources available through the future capable of Orientalism to the extent that ou
... Show MoreThis article studied some linear and nonlinear optical characteristics of different pH solutions from anthocyanin dye extract at 180 oC from red cabbage. First, the linear spectral characteristics, including absorption and transmittance in the range 400-800 nm for anthocyanin solution 5% v/v with different pHs, were achieved utilizing a UV/VIS spectrophotometer. The experimental results reveal a shift in the absorption toward the longer wavelength direction as pH values increment. Then, the nonlinear features were measured using the Z-scan technique with a CW 532 nm laser to measure the nonlinear absorption coefficient through an open aperture. A close aperture (diameter 2 mm) calculates the nonlinear refractive index. The open Z-scan sh
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