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.
In this study, stabilization of expansive soils using waste materials namely; Cement Kiln Dust (CKD), and waste plastic bottles (WPB) was experimentally investigated. Using CKD and WPB are exponentially increasing day by day, due to their capability to solve both environmental and geotechnical problems successfully. Expansive soils were collected from locations with a wide range of plasticity index (PI) (15 - 27) and liquid limit (LL) (35% - 64%). Stabilizer percentages were varied from 0% to 20%, and curing durations for CKD cases were 7 and 28 days. Results showed the best percentages of CKD and WPB are 12% of each one respectively. LL, plastic limit (PL), and swelling percent (SP) loss were observed, which are 46%, 55%, and 96% respec
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Background: Ultrasonography has been used to examine the thickness of the lower uterine segment in women with previous cesarean sections in an attempt to predict the risk of scar dehiscence during subsequent pregnancy. The predictive value of such measurement has not been adequately assessed. Objectives: To correlate lower uterine segment thickness measured by trans abdominal ultrasound in pregnant women with previous cesarean section with that measured during cesarean section by caliper and to find out minimum lower uterine segment thickness indicative of integrity of the scar.Methods: A prospective observational study at Elwyia Maternity Teaching Hospital, from January 2011 to January 2012. A total of 143 women were enrolled in the stu
... Show MoreThe purpose of this research is to identify heritage and highlight its value by drawing on its decorative vocabulary and integrating it with the Arabic calligraphy to revive the heritage in a contemporary style, and to create decorative design units inspired by Sadou and Arabic calligraphy and to employ them in the modern fashion designs. The applied descriptive approach has been used in this research, and the research community is made of women in Riyadh area. The tools used in this research were the questionnaire and the observation. The most important results of the research are: the design of decorative units from the integration of Arabic calligraphy and the decorations of the Saduo, and the use of these units in the design of the o
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The research aims to identify the magnitude of the impact of external debt on the gross domestic product in Morocco, and the importance of research lies in the role that external debt plays in addressing structural imbalances, if it is best disposed of according to well-studied economic plans by specialists in this regard, especially if these debts are directed with Other resources, as it helps pay the costs of these debts (debt servicing) that the external debt also raises the level of gross domestic product, and the research starts from the hypothesis that: There is an effect of foreign debt on the GDP in Morocco, has contributed in one way or another to The exacerbation of the external debt, which affected the m
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The paper deals with the marked vocabulary of Russian and Arabic language, and the extrapolated to the phraseological layer of the mentioned language systems. Specificity of the functioning of this process is presented against the backdrop of the peculiarities of the existence of Russian and Arabic languages. Attention is focused on the fact that linguistic markers should be considered as a kind of keys that represent the specificity of the experience of being experienced by an individual in ontological reality. It is asserted that marking can be revealed practically at all levels of the language polysystem, but it is especially productive on its lexical layer, in particular, on the basis of lexicology and ph
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A new Schiff bases ligand 4- ((2-hyolroxy phenylimino) methyl) -2, 6-dimethoxyphenol derived from condensation of 2- amino phenol with 4-hydroxy -3, 5-dimethoxy benzaldehyde have been synthesized and characterized by spectroscopy, spectra, Mass spectrum and elemental microanalysis (C.H.N). Metal Complexes with ions have been also synthesized and characterized spectroscopic methods spectroscopy, flame atomic absorption, molar conductivity measurements and magnetic susceptibility. These studies indicate that the moler ratio for the complexes. The complexes showed characteristics octahedral geometry with the (O,N) ligand coordinated in bidentate mode while with showed square planer. The enzyme activity of the ligand and i
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This paper investigates the performance evaluation of two state feedback controllers, Pole Placement (PP) and Linear Quadratic Regulator (LQR). The two controllers are designed for a Mass-Spring-Damper (MSD) system found in numerous applications to stabilize the MSD system performance and minimize the position tracking error of the system output. The state space model of the MSD system is first developed. Then, two meta-heuristic optimizations, Simulated Annealing (SA) optimization and Ant Colony (AC) optimization are utilized to optimize feedback gains matrix K of the PP and the weighting matrices Q and R of the LQR to make the MSD system reach stabilization and reduce the oscillation of the response. The Matlab softwar
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In study of effective bioactive compounds, we have synthesized the Co((ІІ), Mn(ІІ), Fe(ІІ), Cu(ІІ), Ni(ІІ), and Zn(ІІ) complexes of the Schiff base derived from trimethoprim and2'-amino-4-chlorobenzophenone and characterized by spectroscopic (NMR, IR, Mass, UV–vis,), analytical, TGA studies and magnetic data .The solution electronic spectral study suggests the stoichiometry of the synthesized complexes and Elemental analysis detected the square planer and octahedral geometry of the compounds. The prepared metal complexes presented promoted efficiency versus the screened bacterial (Escherichia Coli and Staphylococcus aureus) antibacterial efficacy against (Staphylococcus aureus, Salmonella spp., E. coli, Vibrio spp., Pseud
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