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.
في هذا البحث تم تحضير المركبات المعدنية الجديدة لأيونات البلاتين (الرباعي) و الذهب (الثلاثي) مع ليكاند قاعدة مانخ جديد مشتق من السيبروفلوكساسين . تم استخدام المعقدات بعد ذلك كمصدر لتحضير جزيئات عن طريق ترسيب المعقدات على مسام دقائق السيليكا النانوية. Si/Au2O3 Si/PtO2 تم تشخيص الليكاند و معقداته
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This study expands the state of the art in studies that assess torsional retrofit of reinforced concrete (RC) multi-cell box girders with carbon fiber reinforced polymer (CFRP) strips. The torsional behavior of non-damaged and pre-damaged RC multi-cell box girder specimens externally retrofitted by CFRP strips was investigated through a series of laboratory experiments. It was found that retrofitting the pre-damaged specimens with CFRP strips increased the ultimate torsional capacity by more than 50% as compared to the un-damaged specimens subjected to equivalent retrofitting. This indicated that the retrofit has been less effective for the girder specimen that did not develop distortion beforehand as a result of pre-loading. From
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This study expands the state of the art in studies that assess torsional retrofit of reinforced concrete (RC) multi-cell box girders with carbon fiber reinforced polymer (CFRP) strips. The torsional behavior of non-damaged and pre-damaged RC multi-cell box girder specimens externally retrofitted by CFRP strips was investigated through a series of laboratory experiments. It was found that retrofitting the pre-damaged specimens with CFRP strips increased the ultimate torsional capacity by more than 50% as compared to the un-damaged specimens subjected to equivalent retrofitting. This indicated that the retrofit has been less effective for the girder specimen that did not develop distortion beforehand as a result of pre-loading. From
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Amaranthus viridis L. belongs to the Amaranthaceae family. It is a rich source of numerous phytochemicals and amino acids. The objective of this work was to optimize Ultrasound-Assisted Extraction (UAE) based on the extraction yield and Thin-Layer Chromatography (TLC) profile under different conditions, to compare the optimized UAE to the Soxhlet extraction method and evaluate the cytotoxic effects of the ethyl acetate fraction of the 80 % ethanolic extract on the SKGT-4 (human esophageal adenocarcinoma), AGS (human gastric adenocarcinoma) and A431 (human epidermoid carcinoma). A one-factor at a time experiment was carefully designed to assess the influence of the following factors on the extraction: time, frequency, solid-to-solven
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Semi-parametric models analysis is one of the most interesting subjects in recent studies due to give an efficient model estimation. The problem when the response variable has one of two values either 0 ( no response) or one – with response which is called the logistic regression model.
We compare two methods Bayesian and . Then the results were compared using MSe criteria.
A simulation had been used to study the empirical behavior for the Logistic model , with different sample sizes and variances. The results using represent that the Bayesian method is better than the at small samples sizes.
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We have investigated twenty five patients with type-2 diabetes mellitus aged (35-60) years and fifteen healthy persons as control group to detect Anti-Helicobacter pylori IgG antibody. All studied groups were carried out to measure fasting blood sugar, anti- Glutamic acid decarboxylase (GAD), anti-? islets cells antibody by IFAT, Anti-H. pylori IgG antibody by ELISA technique. There was significant elevation in the concentration of fasting blood sugar than in control group (P < 0.05), the patients had negative results for anti-GAD antibody and anti- ? islets cells antibody, there were significant differences (P < 0.05) of anti-H. pylori IgG antibody in 28 % of patients had type-2 diabetes than control group. This lead to suggestion that typ
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Public budget is the government's tool in achieving the objectives of economic and social development is the accounting curriculum to estimate state revenues and expenditures for years to come, as well as to have legal status as it is after the adoption of an official statement of government units to spend funds on items planned at the same time is a statement collect resources to finance these appropriations, if the primary objective of the budget initially limited to the achievement of financial and legislative control have evolved this function in the area of public administration turned attention from mere imposition of control over the money, which provides information to assist the Department to utilize available resources and prog
... Show MoreThe mucilage was isolated from mustard seeds and identification by some different methods like, thermo gravimetric, FTlR., X-ray powdered, proton NMR, FTIR spectra of the three gums contain different functional group in the gums, major peaks bands noticed were belong to OH (3410.15 – 3010.88) group from hydroxyl group, CH aliphatic (2925-2343.51), C-O (1072.42-1060.85) group and C=O 1743.65, Thermo chemical parameters of mucilage was evaluated and compared with the standard gums, Results indicated the mucilage was decomposed in 392°C and mass loss 55%, The X ray process found the mucilage had single not sharp peak
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