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 study aims at knowing the actual uses of instructional technology in teaching history subject in the colleges of education for humanities from the college staff members' perspectives and their attitudes towards it. The sample of the study consisted of (24) instructors from the Colleges of Education for the Humanities, the College of Basic Education- Haditha and the College of Education for Women, the study used the descriptive method, and the questionnaire was consisted of (50) items, and the psychometric properties of the instrument of the study were extracted. The researchers used the appropriate statistical means to analyze the data, and the results of the study showed the following: the teaching staff attitudes towards the use of
... Show MoreThe lead has adverse effects in contamination the aquatic environment, for this reason, a laboratory simulation was conducted using kaolinite collected from the Ga’ara Formation at western Iraq to be considered as a natural sorbent material that can be addressed Pb2+ from the aqueous environments. The Energy-Dispersive X-ray Spectroscopy and atomic absorption spectroscopy clarifying very fine grains and pure phase with a very little quantity of quartz and has a number of active sites for adsorption. The sorption of kaolinite for the Pb2+ has been carefully tested by several designed laboratory experiments. Five lead solutions of different concentrations (25, 50, 75, 100 and 125 ppm) were tested under different values of pH (1.3-9)
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Freedom of opinion is one of the freedoms that constitutions and laws have guaranteed to protect and is exercised through the media. However, this freedom must be exercised within certain limits, which include respecting a person’s right to preserve his privacy, image and reputation. Just as the media has the freedom to exercise its right to publish and express its opinion with complete frankness, it is in return obligated to respect the rights of others and avoid defaming them and violating their privacy. If the media does this, the injured party has the right to demand compensation for the damage he has suffered as a result.
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Urine specimen (253 samples) had been collected from urinary tract infections. The study showed that Proteus mirabilis was responsible for (11.85%) of the urinary tract infections. Also, the study had declared that the ratio of isolation of this bacterium from women was (7.51%) and it was higher than the ratio in both men and children which ranged (1.58%) and (2.76%) respectively. Morphological and biochemical studies had been applied to characterize this bacterium as well as other kinds of microorganisms that were isolated from urinary tract infection in this study. The study deals with typing methods such as using biotyping and typing according to Dienes phenomenon beside the succestibility to antibiotics. The results had shown that the s
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KHalaf Alhmar and what we get from his poem
Background: Birth defects are the leading cause of both neonatal and post-neonatal deaths, as an estimated 240,000 neonates die in their first month of life worldwide each year. In Iraq, local studies have shown varying frequencies and types of congenital anomalies. Objectives: To provide new insight into the incidence and types of congenital anomalies and to explore their possible risk factors in Baghdad City. Methods: In this cross-sectional observational study, a total of 2007 neonates were enrolled from six hospitals in Baghdad during the period extending between September and December 2020. Hospital records and personal interviews were used for data collection. These data included the neonates' demographic and clinical characte
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Tennessee Williams' The Glass Menagerie is peopled with powerless and defenseless characters. These characters are hypersensitive and in a constant search for protection. They are highly pressurized by life and its bitter reality. Adding to that they are haunted by their fear from the future. They are escapist who try their best to hide behind a protective shield. Their tireless search for security is never smothering. Their only possible defensive mechanism is to flee from their immediate hostile and aggressive surrounding. To achieve this aim, each character has improvised a method that suits him / her best. For example, one of the characters (Tom) sought refuge in alcohol and late night shows. While the other (Amanda) chose to re-live th
... Show MoreThe ground state proton, neutron and matter densities andcorresponding root mean square radii of unstable proton-rich 17Neand 27P exotic nuclei are studied via the framework of the twofrequencyshell model. The single particle harmonic oscillator wavefunctions are used in this model with two different oscillator sizeparameters core b and halo , b the former for the core (inner) orbitswhereas the latter for the halo (outer) orbits. Shell model calculationsfor core nucleons and for outer (halo) nucleons in exotic nuclei areperformed individually via the computer code OXBASH. Halostructure of 17Ne and 27P nuclei is confirmed. It is found that thestructure of 17Ne and 27P nuclei have 25 / 2 (1d ) and 1/ 2 2s -dominantconfigurations, resp
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