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 nuclear charge density distributions, form factors andcorresponding proton, charge, neutron, and matter root mean squareradii for stable 4He, 12C, and 16O nuclei have been calculated usingsingle-particle radial wave functions of Woods-Saxon potential andharmonic-oscillator potential for comparison. The calculations for theground charge density distributions using the Woods-Saxon potentialshow good agreement with experimental data for 4He nucleus whilethe results for 12C and 16O nuclei are better in harmonic-oscillatorpotential. The calculated elastic charge form factors in Woods-Saxonpotential are better than the results of harmonic-oscillator potential.Finally, the calculated root mean square radii usingWoods-Saxonpotentials ho
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One of the Kurdish scholars who have research on the doctrinal issues of the author of this brief message that we are about to achieve; to bring it out better and in order to preserve the Islamic heritage, and revive what can be revived after being close to death by staying in libraries here and there, out of reach of hands, carrying dust Years of forgetting and marginalization, and this letter is the papers in the door of the divorce entitled his author "Iqdar al-dream for those who did not sign divorce commoner," and the name is evident the name, and the intention of the author through writing, where he wrote it to answer some of the knowledgeable among the public at the time So they decreed that there was no divorce from commoners D w
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The research revolves around the collection of pending hadiths on Abdullah bin Amr bin Al-Aas, may God be pleased with them, and it is required in these hadiths that they be among those in which there is no room for diligence, such as telling about unseen matters, signs of the Hour, or a statement of virtue and reward for obedience, or punishment for disobedience.
The research consists of an introduction, two topics, and a conclusion of the first topic in the translation of Abdullah bin Amr bin Al-Aas, and the second topic, the pending hadiths of Abdullah bin Amr bin Al-Aas, may God be pleased with them, in which there is no room for diligence in collecting and studying
At the end of this rese
The research dealt with the design of the cost accounting system for the transport service and its Role in improving the efficiency of pricing decisions through the application of the cost system based on ABC activities. The main activities were defined and cost guides were to measure the cost of each service and to determine the cost of each service for the purpose of providing management with appropriate information and pricing decisions The problem of research in the lack of adoption by some public companies in the service sector on the cost accounting system to calculate the cost of service as well as the lack of identification of productive activities and service activities and therefore cannot make the appropriate decision t
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Mammals are under threat worldwide due to deforestation, hunting, and other human activities. In Iraq, a total of 93 species of wild mammals have been recorded including species with global conservation concern. Bamo Mountain is situated within the Zagros Mountains in northern Iraq which is a suitable habitat for wild mammals. Due to scarcity of the field survey efforts and cryptic behavior, monitoring of the wild mammals fauna in Zagros Mountain seems challenging. Therefore, we used a camera trap which seems to be an ideal way to determine species diversity of wild mammals in Bamo Mountain. Moreover, interviews with local villagers were performed. The mammalian diversity of Bamo Mountain is not fully explored but seemed threatened by lo
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Darcy-Weisbach (D-W) is a typical resistance equation in pressured flow; however, some academics and engineers prefer Hazen-Williams (H-W) for assessing water distribution networks. The main difference is that the (D-W) friction factor changes with the Reynolds number, while the (H-W) coefficient is a constant value for a certain material. This study uses WaterGEMS CONNECT Edition update 1 to find an empirical relation between the (H-W) and (H-W) equations for two 400 mm and 500 mm pipe systems. The hydraulic model was done, and two scenarios were applied by changing the (H-W) coefficient to show the difference in results of head loss. The results showed a strong relationship between both equations with correlation coefficients of 0.999,
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Abstract
The research’s goal lies in demonstrating the impact of the Federal Financial Supervision Endowment through the process of auditing the performance of the entities subject to its audit as to improve the performance of these entities, especially if the performance audit method is one of the newly applied methods that are compatible with the standards issued by the International Organization of Financial Supervision and Accounting Institutions which is the method of auditing performance according to the performance evaluation guide for programs and policies issued by the Federal Office of Financial Supervision.
T
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