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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God Almighty put in his great book secrets that do not end, and wonders that do not expire, for he is the one from which the scholars are not satisfied, and he does not create due to the multitude of response, and it is the comprehensive and inhibitory book that God conceals to the worlds, and he challenged the two heavyweights to come up with something like it.
At all times, issues arise in the Noble Qur’an that fit the needs of the people of that time and their culture, for it is an eternal book, characterized by the ability to give, extend and respond to addressing the problems of the age and its variables, when the Arabs had little luck at the time of the message’s descent from the scientific culture, and their proficienc
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Abstract
That Iraq's dependence on the revenues of the oil product in financing its development programs and growth rates , Making the economy affected by external forces represented by fluctuations in crude oil prices in the global market, Which is directly reflected on the performance and efficiency of the Iraqi economy.
The study adopted its objectives to analyze the time series for the period (1988 - 2015) through the use of standard and statistical methods, Four standard models were estimated to reach those targets, Where the results of the stability test showed instability of most variables at their original level, But to achieve stability when taking the first differences, While the result
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Abstract
Objectives: To find out the association between enhancing learning needs and demographic characteristic of (gender, education level and age).
Methods: This study was conducted on purposive sample was selected to obtain representative and accurate data consisting of (90) patients who are in a peroid of recovering from myocardial infarction at Missan Center for Cardiac Diseases and Surgery, (10) patients were excluded for the pilot study, Data were analyzed using descriptive statistical data analysis approach of frequency, percentage, and analysis of variance (ANOVA).
Results: The study finding shows, there was sign
... Show MoreThe research aims to identify the effect of the training program that is based on integrating futuristic thinking skills with classroom interaction patterns on mathematics teachers in order to provide their students with creative solution skills. The research sample consisted of 31teachers (15 teachers for the experimental group and 16 for the control groups). The researcher developed a measure for the academic self-efficacy consisting of (39) items. Its validity, reliability, coefficient of difficulty and discriminatory power were estimated. To analyze the findings, the researcher adopted the Mann-Whitney (U) test and the effect size, and the findings were as follows: There is a statistically significant difference at the significance leve
... Show MoreBackground: Type 2 diabetes mellitus is a condition characterized by an elevation of oxidative stress, which has been implicated in diabetic progression and its vascular complications. Aim: Assessing the impact of gliclazide modified release (MR) versus glimepiride on oxidative stress markers, glycemic indices, lipid profile, and estimated glomerular filtration rate in uncontrolled type 2 diabetic patients on metformin monotherapy. Methods: This was an observational comparative study conducted in Thi-Qar specialized diabetic, endocrine, and metabolism center. Sixty-six patients were randomized into two groups based on the addition of the sulfonylureas (SUs). Group 1 (33 patients) was on gliclazide MR, whereas Group 2 (33 patients)
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Abstract:
The aerial part of Ephedra foliata Family Ephedraceae have long been used in traditional medicine and now Ephedra species have medicinal, ecological, and commercial value. The variety of pharmacological actions of this plant is due to its chemical constituents. Ephedrine and
related alkaloids; are the newly potential medicinal value of Ephedra supplements for weight loss or performance improvement. Other pharmacological actions like antibacterial and antifungal effects of the phenolic acid compounds, the immunosuppressive action of the polysaccharides, and the antitumor action of flavonoids. The genus of this plant wildly distributed t
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The Vulnerable Indian Roofed Turtle Pangshura tecta (Gray, 1831) (Testudines: Geoemydidae) occurs in the Sub-Himalayan lowlands of India, Nepal, Bangladesh, and Pakistan. Little is known about its natural history, no studies have been conducted revealing its natural predators. In this study, a group of Large-billed Crow Corvus macrorhynchos Wagler, 1827 (Passeriformes: Corvidae) was observed hunting and predating on an Indian Roofed Turtle carcass in the bank of river Kuakhai, Bhubaneswar, India. The first record of this predation behaviour is reported and substantiated by photographic evidence.
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Abstract
This Research aims for harnessing critical and innovative thinking approaches besides innovative problem solving tools in pursuing continual quality improvement initiatives for the benefit of achieving operations results effectively in water treatment plants in Baghdad Water Authority. Case study has been used in fulfilling this research in the sadr city water treatment plant, which was chosen as a study sample as it facilitates describing and analyzing its current operational situation, collecting and analyzing its own data, in order to get its own desired improvement opportunity be done. Many statistical means and visual thinking promoting methods has been used to fulfill research task.
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