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.
New chelating ligand derived from triazole and its complexes with metal ions Rhodium, Platinum and Gold were synthesized. Through a copper (I)-catalyzed click reaction, the ligand produced 1,3-dipolar cycloaddition between 2,6-bis((prop-2-yn-1-yloxy) methyl) pyridine and 1-azidododecane. All structures of these new compounds were rigorously characterized in the solid state using spectroscopic techniques like: 1HNMR, 13CNMR, Uv-Vis, FTIR, metal and elemental analyses, magnetic susceptibility and conductivity measurements at room temperature, it was found that the ligand acts as a penta and tetradentate chelate through N3O2, N2O2, and the geometry of the new complexes are identified as octahedral for (Rh & Pt) complexes a
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novel spectrofluorimetric flow injection analysis (FIA) method was developed for the selective quantification of ascorbic acid via fluorescence quenching of serotonin hydrochloride. The system employs a custom-designed photometric array comprising 16 irradiation sources arranged in a dual-axis matrix—eight aligned horizontally and eight orthogonally, enabling multi-angle excitation and enhanced spectral resolution. Fluorescence signals were captured using a twin-pair solar cell detector, offering high sensitivity and minimal optical interference. The method exhibited a linear calibration range of 0.1–30 limit of detection (LOD) of 0.025 μ μ g/mL with a correlation coefficient (r mol /L, equivalent to 4.403 * 10 4 μ 2 ) of 0.9966, a g
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Some microorganisms can produce nanocellulose, which is known as bacterial nanocellulose (BNC); the most active bacterial producer is acetic acid bacteria (AAB), which is a gram-negative, motile and obligate aerobic belongs to the family Acetobacteraceae. Bacterial nanocellulose has excellent attention in medical (surgical domain), industrial and pharmaceutical fields because of its flexible properties, characteristics and advantages. So, in this study, the AAB (5AC) isolate was isolated from apple vinegar. The production of BNC was performed by using a natural medium called palm dates liquid medium, the produced bacterial Cellulose was purified by using the sodium hydroxide method; it was observed that the wet weight of the BNC was a
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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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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 MoreColonialism as a movement was very popular in Europe more than two centuries before. It aimed at controlling and exploiting several countries in Africa and Asia in addition to imposing their power and control on uninhabited islands. It received adherence and criticism as well. There also appeared activists and nations who stood against it and its practices. English novels discussed this notion greatly by pointing out the bad practices of the colonizers and how the colonized received them. This paper explores two narrative fictions that tackle the different aspects of the term. While Defoe, in Robinson Crusoe (1719), shows a colonial European figure who expresses his superiority, Wells, in “The Country of the Blind” (1904), deconstructs
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Aspartate aminotransferase was purified from urine and serum of patients with type 2 diabetes in a 2 steps procedure involving dialysis bag and sephadex G-25 gel filtration (column chromatography). The enzyme was purified 346.23 fold with 1467% yield and 3.46 fold with 142.85% yield in urine and serum of patients with type 2 diabetes respectively. The purified enzyme showed single peak. The results of this study revealed that AST activity of type 2 diabetes urine and serum increased significantly (p<0.001) compared with control group.
The artificial silk (Rayon) was produced from the fronds of date palms which was taken from date palm trees (type Al-Zahdi) from the Iraqi gardens. Two main parts of the frond, namely leaves and stalks were used in this study to produce rayon. The palm fronds were converted into a powder of 90-180 micrometers. Major steps were used to produce rayon; delignification, bleaching and finally dissolution. Modified organosolv method which uses organic solvent method was applied to remove high lignin content. Three variables were studied in the delignification process: temperature, the ratio of ethanol to water and digestion time. The results showed that the best percent of lignin removal was (97%) which occured at; digestion time (80 minutes), te
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