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.
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This research utilized natural asphalt (NA) deposits from sulfur springs in western Iraq. Laboratory tests were conducted to evaluate the performance of an asphalt mixture incorporating NA and verify its suitability for local pavement applications. To achieve this, a combination of two types of NA, namely soft SNA and hard HNA, was blended to create a binder known as Type HSNA. The resulting HSNA exhibited a penetration grade that adhered to Iraqi specifications. Various percentages of NA (20%, 40%, 60%, and 80%) were added to petroleum asphalt. The findings revealed enhanced physical properties of HSNA, which also satisfied the requirements outlined in the Iraqi specifications for asphalt cement. Consequently, HSNA can serve as an
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This research includes a study of the ability of Iraqi porcelanite rocks powder to remove the basic Safranine dye from its aqueous process by adsorption. The experiments were carried out at 298Kelvin in order to determine the effect of the starting concentration for Safranin dye, mixing time, pH, and the effect of ionic Strength. The good conditions were perfect for safranine dye adsorption was performed when0.0200g from that adsorbed particles and the removal max percentage was found be 96.86% at 9 mg/L , 20 minutes adsorption time and at PH=8 and in 298 K. The isothermal equilibrum stoichiometric adsorption confirmed, the process data were examined by Langmuir, Freundlich and Temkin adsorption equations at different temperatures
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The present study aimed to identify the therapeutic evaluation of chitosan extracted from the fungus cushroom and pure chitosan on glucose and lipid profile in the blood of 35 male rabbits with hyperlipidemia induced experimentally by cholesterol. The tests included estimation of glucose levels, total cholesterol, triglycerides, high-density lipoproteins, low-density lipoproteins, and very low-density lipoproteins. hyperlipidemia was induced in the male rabbits used in the study which was administered orally with cholesterol 150mg/kg body weight for a week. rabbits were divided into seven groups: control, cholesterol, pure chitosan, mushroom chitosan, cholesterol and pure chitosan, cholesterol and mushroom chitosan and cholestero
... Show MoreThis study was carried out to investigate the cytogenetic effects of crude aqueous extract of Lycium barbarum on the roots tip of Allium cepa Using three concentration 125, 25, 50 mg/ ml for 2, 4, 6 hours treatment periods.This study were included some cytogenetic analysis such as mitotic index , phase index and chromosome aberration. The data showed that the treatment with 50mg/ml for 6huors led to reduce the mitotic index less than 50% . This reduction considered to have toxic and sublethal effect . These results revealed mutagenic potency by inducing differents type of chromosome aberration.
جريت دراسة مختبرية لمعرفة تأثير الزيت الطيار لقشور ثمار نبات النارنج الصفرC. aurantium تجاه النمو السطحي للفطريات Penicillium expansum، Aspergillus flavus و Fusarium oxysporum ، أظهرت نتائج الفعالية التثبيطية للزيت الطيار تأثيراً معنويا متفاوتاً في الفطريات المشمولة بالدراسة، إذ كان الزيت الطيار أكثر تأثيرأَ في الفطر P. expansum تلاه الفطر A. flavus ،في حين كان الفطر oxysporum F.أقل حساسية تجاه الزيت الطيار. بصورة عامة اظهر الزيت الطيار تأثيرا تثبيطيا
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This study aims at identifying the impact of the application of IFRS 15 "Revenue from contracts with customers on the quality of financial reporting, through application to faculty members in the accounting departments of Iraqi universities and auditors. The problem of the study was the multiplicity of accounting rules and standards Which deals with the issues of revenue recognition , as well as the lack of consistency of most of them with the common framework of financial accounting, which results in low quality of financial reporting in the current financial statements, where the formulation of one hypothesis was the lack of relationship of significant significance The application of IFRS 15 "Recognition of rev
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