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.
This research involves the application of spectral indices and GIS techniques coupled with the Analytic Hierarchy Process, to assess Baglia site, eastern Missan, as a water harvesting potential site. The AHP and pairwise comparison have been used through select four criteria including TWI, TRI, MNDWI, and NDSI, which were deemed as impact factors for this study. All these criteria have been weighted according to their significance in the water harvesting system. The findings of the AHP analysis method explained that the WH potential zones in Baglia site were divided into three zones, namely, high, medium, and low suitability. The findings demonstrate that Bglia site, where the highly appropriate zone is located in the deep v
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Fluorescent proteins (FPs) have revolutionised the life sciences, but the chromophore maturation mechanism is still not fully understood. Here we photochemically trap maturation at a crucial stage and structurally characterise the intermediate.
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The current study was conducted to determine the effect of crude oil on the pigments and protein content of Hydrilla verticillata plant from December 2018 to February 2019. Hydrilla verticillata was exposed to different concentrations of treatments (0.01,0.05,0.10,0.20) % of crude oil for 24 days. The pigments content was evaluated as total chlorophyll, a, b, carotene and protein content within plant tissues during the days (1, 8, 16 and 24).
The results of this study showed an increment in the total chlorophyll values of plant, which is exposed to different concentrations of crude oil compared to the control treatment during the first day of the experiment and decreased compared to cont
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Optoelectronic devices, widely used in high energy and nuclear physics applications, suffer severe radiation damage that leads to degradations in its efficiency. In this paper, the influence of gamma radiation (137Ce source) and beta radiation (90Sr source) on the photoelectric parameters of the Si solar cell, based on the I–V characterization at different irradiation exposer, has been studied. The penetrating radiation produces defects in the base material, may be activated during its lifetime, becoming traps for electron–hole pairs produced optically and, this will, decrease the efficiency of the solar cell. The main objective of the paper is to study and measure changes in the I–V characteristics of solar cells, such as efficienc
... Show MoreSeveral observations have showed the synergistic effect of nutrients elements and phytoestrogens on bone resorption elimination. Zn is one of the trace elements found to increase the stimulatory effect of phytoestrogens including genestein, coumestrol and daidzein on bone formation; however the synergistic modulation of Zn, genestein, coumestrol and daidzein on proinflammatory-producing T-cells and receptor activator of NFκB ligand (RANKL) expression that implicated in osteoclast formation is still open area to debate. This study found that Zn enhanced the inhibitory effect of genistein, daidzein and coumestrol on TNF-α expression; however the same effect was shown with daidzein on IL-1β expression while there is no furthe
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Attempts were made over the years to achieve economic and easy methods for water purification. This could well save time, cost, and earn a good process quality for many countries. This study aims to enhance the purification process parameters for Congo red dye polluted water and introduce a developed methodology with an impact on many associated parameters such as the time for water purification process. The research proposes a method to achieve this time reduction by mixing gold nanoparticle (AuNPs) (prepared by chemical reduction method) with Titanium dioxide nanoparticles (TiO2NPs) (prepared by sol-gel techniques). The resulting mixture is incorporated into PVA host to synthesize a hard disk used as a purification d
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The theory of general topology view for continuous mappings is general version and is applied for topological graph theory. Separation axioms can be regard as tools for distinguishing objects in information systems. Rough theory is one of map the topology to uncertainty. The aim of this work is to presented graph, continuity, separation properties and rough set to put a new approaches for uncertainty. For the introduce of various levels of approximations, we introduce several levels of continuity and separation axioms on graphs in Gm-closure approximation spaces.
The purpose of this study was to determine the impact of added soy protein humidifier concentration of different ratios of bovine Alberkr mix that was to bring the old chicken shop pure beef in five ratios