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.
This research examines how Al-Sabah newspaper addressed violations of women’s rights in Iraq. Using content analysis, a sample of 157 articles published from July 1, 2021, to December 31, 2021, was analyzed. The study reveals diverse journalistic treatments, with interpretive approaches employed in addressing violations of Iraqi women’s rights. Social violations of women’s rights were the primary focus, and various forms of newspaper articles, including opinion pieces, were commonly used.
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Three types of extracts ( aquatic, alcoholic, and oily ) were prepared from the fruits of coconuts, and a series of chemical tests were conducted in addition to the use of the FTIR equipment to determine the active locations in the prepared extracts. The results indicated the presence of active compounds (tannins, saponins, flavonoids, turbines and steroids) in the extracts prepared from the fruits of coconuts, also the antimicrobial capability of these extracts were tested on pathogenic bacteria isolated from wounds and burns infections cases. The results proved that the concentration 80 mg/ml of the aquatic extract is the minimum inhibitory concentration for the microbes: Proteus vulgaris and Pseudomonas fluorescence, while the
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A common field development task is the object of the present research by specifying the best location of new horizontal re-entry wells within AB unit of South Rumaila Oil Field. One of the key parameters in the success of a new well is the well location in the reservoir, especially when there are several wells are planned to be drilled from the existing wells. This paper demonstrates an application of neural network with reservoir simulation technique as decision tool. A fully trained predictive artificial feed forward neural network (FFNNW) with efficient selection of horizontal re-entry wells location in AB unit has been carried out with maintaining a reasonable accuracy. Sets of available input data were collected from the exploited g
... Show MoreAchieving goals effectively reflects the success of the institution. However, unless this indicator is coupled with efficiency when achieving goals, the institution will be equal in its achievements, and distinction will remain unachieved. Perhaps the role of the teaching staff in pushing the institution or college towards brilliance focuses on their ability to motivate people on the one hand and their interest in achieving brilliance for the institution. On the other hand, the importance of the research lies in the institution’s reaching a prominent position through the brilliance and creativity of teaching and achieving competition between institutions that make it more brilliance. The study seeks to achieve the goal of the real
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This study was done to find a cheap, available and ecofriendly materials that can remove eosin y dye from aqueous solutions by adsorption in this study, two adsorbent materials were used, the shells of fresh water clam (Cabicula fluminea) and walnut shells. To make a comparison between the two adsorbents, five experiments were conducted. First, the effects of the contact time, here the nut shell removed the dye quickly, while the C. flumina need more contact time to remove the dye. Second, the effects of adsorbent weight were examined. The nut shell was very promising and for all used adsorbent weight, the R% ranged from 94.87 to 99.29. However C. fluminea was less effective in removing the dye with R% ranged from 47.59 to 55.39. The thi
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In this study, the photodegradation of Congo red dye (CR) in aqueous solution was investigated using Au-Pd/TiO2 as photocatalyst. The concentration of dye, dosage of photocatalyst, amount of H2O2, pH of the medium and temperature were examined to find the optimum values of these parameters. It has been found that 28 ppm was the best dye concentration. The optimum amount of photocatalyst was 0.09 g/75 mL of dye solution when the degradation percent was ~ 96 % after irradiation time of 12 hours, while the best amount of hydrogen peroxide was 7μl/75 mL of dye solution at degradation percent ~97 % after irradiation time of 10 hours, whereas pH 5 was the best value to carry out the reaction at the highest deg
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The phenyl hydrazine was react readily with acetic acid chloride in [1:2] ratio in alkyl of ethanolic solution, and refluxe for five hours to produce a new ligand of (N-Carboxymethyl-N-phenyl-hydrazino)-acetic acid [H2L].
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