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.
From the sustainability point of view a combination of using water absorption polymer balls in concrete mix produce from Portland limestone cement (IL) is worth to be perceived. Compressive strength and drying shrinkage behavior for the mixes of concrete prepared by Ordinary Portland Cement (O.P.C) and Portland limestone cement (IL) were investigated in this research. Water absorbent polymer balls (WAPB) are innovative module in producing building materials due to the internal curing which eliminates autogenous shrinkage, enhances the strength at early age, improve the durability, give higher compressive strength at early age, and reduce the effect of insufficient external curing. Polymer balls (WAPB) had been used in the mixes of thi
... Show MoreBackground: A great dental and biomedical interest had been paid to silver nanoparticles because of their antimicrobial activity. Objective: To evaluate the antimicrobial and cytotoxic activity of a newly developed Nano-silver fluoride that was synthesized from moringa oleifera leaf extract against S. mutants. Material and method: The green synthesis method was used to prepare Nano-silver fluoride from moringa oleifera leaf extract. The minimum inhibitory concentration and the minimum bactericidal concentration were evaluated using brain heart infusion plates, while the cytotoxicity was evaluated by the hemolytic activity. Results: Nano-silver fluoride had a bactericidal and bacteriostatic effect (MIC was 60 ppm and MBC was 120 pp
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The cathodic deposition of zinc from simulated chloride wastewater was used to characterize the mass transport properties of a flow-by fixed bed electrochemical reactor composed of vertical stack of stainless steel nets, operated in batch-recycle mode. The electrochemical reactor employed potential value in such a way that the zinc reduction occurred under mass transport control. This potential was determined by hydrodynamic voltammetry using a borate/chloride solution as supporting electrolyte on stainless steel rotating disc electrode. The results indicate that mass transfer coefficient (Km) increases with increasing of flow rate (Q) where .The electrochemical reactor proved to be efficient in removing zinc and was abl
... Show MoreAbstract Sweden is today one of the most active European countries in the regional and international environment despite the adoption of neutrality as a guiding principle in its foreign policy. For more than two centuries, the length of time for Swedish neutrality has made it a global standard, an agreed foreign policy at home and a political culture rooted in institutions and society. Swedish. Although discussions are still underway on Swedish security and foreign policies after the end of the Cold War, especially cooperation with NATO through the Partnership for Peaceprogram, EU accession and its impact on the principle of neutrality in foreign policy. Sweden, however, insists that it still maintains neutrality, but more adequately, in
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Oil pollution of the soil due to a leakage in oil tubes, transportation of products, or during oil excavations can change the soil physical and mechanical, chemical, and biological properties. Consequently, the soil may or may not be eligible for engineering construction projects and it may need a significant treatment. Therefore, it is required to have a better understanding of the general behavior and the corresponding geotechnical properties upon pollution particularly for those areas associated with oil explorations and industry like Thi-Qar Governorate. Fine and coarse soils from two sites at the University of Thi-Qar are artificially contaminated with oil products ranging from 0% to 10% of their dry weight. Testing programs have been
... Show MoreThe antagonism of the rhizospheric bacteria toward pathogenic fungi Macrophomina phaseolina was investigated. Ten soil samples were collected from the rhizospheric zone around Cowpea root (Vignaunguiculata L.). These samples were used as the source of Arbuscular Mycorrhizal fungi (AMF) and rhizobacterial isolates. Twenty-five bacteria were isolated and evaluated as an antagonistic agent against pathogenic fungi. M. phaseolina was isolated from infected roots of Cowpea and used as a pathogen. Twenty-five bacteria were isolated and evaluated as an antagonistic agent against pathogenic fungi. M. phaseolina was isolated from infected roots of Cowpea and used as a pathogen. The synergistic effect between A. siccitolerans and (AMF) Glomusmosseae,
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Periodontitis is a chronic inflammation affecting the tooth-supporting periodontal tissues. It is diagnosed by measuring periodontal parameters. However, documenting this data takes effort and may not discover early periodontitis. Biomarkers may help diagnose and assess periodontitis. This study aimed to evaluate the potential diagnostic of the salivary tumor necrosis factor-α (TNF-α) and receptor-activator of nuclear factor ĸ-B-ligand (RANKL) in distinguishing between periodontitis and healthy periodontium.
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In the present study, activated carbon supported metal oxides was prepared for thiophene removal from model fuel (Thiophene in n-hexane) using adsorptive desulfurization technique. Commercial activated carbon was loaded individually with copper oxide in the form of Cu2O/AC. A comparison of the kinetic and isotherm models of the sorption of thiophene from model fuel was made at different operating conditions including adsorbent dose, initial thiophene concentration and contact time. Various adsorption rate constants and isotherm parameters were calculated. Results indicated that the desulfurization was enhanced when copper was loaded onto activated carbon surface. The highest desulfurization percent for Cu2O/AC and o
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