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 sole aim of the tribe of the Messenger of Allah (PBUH) and his basilica was to revoke the Umayyad heresy and to refute the suspicions through which they climbed to the top of the pyramid of power and legitimized the leadership of Muslims (heresy of hereditary rule) with the sword and blood. Therefore, Imam Hussein (peace be upon him) began his Ashura process with his short divine message in its words and great in its vocabulary and content and even the future of Mohammedan Islam over the times and spaces - what we will see in the folds of research, God willing.
Among a collection of ground beetles from Iraq the new species Acinopus euphraticus was designated and described here. The erection of this new species was mainly built on external features and the description of male genitalia.
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In this study, the circulating fluidized bed was used to remove the Tetracycline from wastewater utilizing a pistachio shell coated with ZnO nanoparticles. Several parameters including, Tetracycline solution flowrate, initial static bed height, Tetracycline initial concentration and airflow rate were systematically examined to show their effect on the breakthrough curve and the required time to reach the adsorption capacity and thus draw the fully saturated curve of the adsorbent. Results showed that using ZnO nanoparticles will increase the adsorbent surface area and pores and as a result the adsorption increased, also the required time for adsorbent saturation increased and thus the removal efficiency may be achieved at mi
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The world is confronted with the twin crisis of fossil fuel depletion and environmental degradation caused by fossil fuel usage. Biodiesel produced from renewable feedstocks such as Jatropha seed oil or animal fats by transesterification offers a solution. Although biodiesel has been produced from various vegetable oils such as Jatropha seed oil, the reaction kinetics studies are very few in literature, hence the need for this study. Jatropha curcas seed oil was extracted and analyzed to determine its free fatty acid and fatty acid composition. The oil was transesterified with methanol at a molar ratio of methanol to oil 8:1, using 1% sodium hydroxide catalyst, at different temperature
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New technologies have risen into popularity causing the Liquid membrane techniques to evolve over other separation techniques due to its high selectivity and recovery, increased fluxes, and reduced investment and operating cost. This work focuses on extracting Methylene Blue (MB), a cationic dye using a simple BLM separation technique from its aqueous phase. It combines extraction and stripping in a single unit operation. The feed phase was an aqueous solution of MB, the solvent chosen was soybean oil for the liquid/organic membrane phase, and tri-octyl amine acted as a carrier. The strip phase was a hydrochloric acid solution for this study. A two-phase equilibrium study was done to choose the correct solvent, carrier,
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The aim of the research is to identify the losses resulting from the terrorist operations and then find a proposed accounting treatment for the losses resulting from the terrorist operations and to indicate their impact on disclosure in the financial statements by reviewing the international standards and local rules and the unified accounting system and not dealing with these losses, Of the financial statements and therefore adversely affect the accounting disclosure as well as the weak commitment of economic units to apply the requirements of accounting measurement and disclosure of losses of terrorist operations in a manner consistent with local and international standards to achieve the Reliability in the financial statement.
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Abstract This research investigates how activated carbon (AC) was synthesized from potato peel waste (PPW). Different ACs were synthesized under the atmosphere's conditions during carbonation via two activation methods: first, chemical activation, and second, carbon dioxide-physical activation. The influence of the drying period on the preparation of the precursor and the methods of activation were investigated. The specific surface area and pore volume of the activated carbon were estimated using the Brunauer–Emmett–Teller method. The AC produced using physical activation had a surface area as high as 1210 m2/g with a pore volume of 0.37 cm3/g, whereas the chemical activation had a surface area of 1210 m2/g with a pore volume of 0.34 c
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Animal fats are a good, promising and ethical alternative source for biodiesel production, but they need more complex treatments than vegetable oils. Iraqi butchery plants waste fats (sheep fat) which are suggested as feedstock to produce biodiesel. This type of fat contains a large quantity of free fatty acids (FFAs) (acid number 49.13 mg KOH/g of fat). The direct transesterification of such fats produce high amount of soap instead of desired biodiesel, so a pre-treatment step (to reduce FFAs) is necessary before transesterification. This step was done by esterification of the free fatty acids in the fat by adding ethanol and using 1% acid catalyst (H2SO4) for 30 minutes. The results showed that the acid number of sheep fat after pre-tr
... Show MoreAniera desert/cola was found new to science and to the Iraqi fauna. The description was
mainly based on external features and male genit