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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Natural convection in a trapezoidal enclosure with partial heating from below and symmetrical cooling from the sides has been investigated numerically. The heating is simulated by a centrally located heat source on the bottom wall, and four different values of the dimensionless heat source length, 1/5, 2/5, 3/5, 4/5 are considered. The laminar flow field is analyzed numerically by solving the steady, two-dimensional incompressible Navier-Stokes and energy equations. The Cartesian velocity components and pressure on a collocated (non-staggered) grid are used as dependent variables in the momentum equations discretized by finite volume method; body fitted coordinates are used to represent the trapezoidal enclosure, and grid generatio
... Show MoreAbstract The present work included morphological, anatomical, and palynological characters for the new species Acaalypha australis L. specimens, which belong to the family Euphorbiaceae. The species recorded in the study for the first time in Iraq. The plants of this species are annual herbs with green, striated or sub – polygonal stem, and branched near bases, Leaves are simple spirally alternate and lanceolate in shape. Flowers are unisexual, arranged in the axial of distinct leafy and cordate bracts, female flower arranged at the bracts bases and each flower with trileafed perianth and superior ovary with trilobed stylar stigma which has dense and coiled stigmatic hairs. Male flowers are arranged as a mixed verticellate inflorescence a
... Show MoreThis work focuses on the use of biologically produced activated carbon for improving the physi-co-chemical properties of water samples obtained from the Tigris River. An eco-friendly and low-cost activated carbon was prepared from the Alhagi plant using potassium hydroxide (KOH) as an impregnation agent. The prepared activated carbon was characterised using Fourier-transform infrared spectroscopy to determine the functional groups that exist on the raw material (Alhagi plant) and Alhagi activated carbon (AAC). Scanning electron microscope–energy-dispersive X-ray spectroscope was also used to investigate the surface shape and the elements that compose the powder. Brunauer–Emmett–Teller surface area analysis was used to evaluate the spe
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134 samples of plants and animals wastes were taken from three different regions outside Baghdad and three different regions in Baghdad. 24 cellulolytic isolates fungi AO, C1, TH1, AN1, R1, TV, PG, AF, B1, L1, AP, TH, AP1, AN3, AO2, A, A1, C, F, AO1, C2, F1, CL and AP2 independent were chosen out of 48 selected fungi. The best optimal conditions for growth were 30ºC and pH 7. The isolates were identified and screened according to the colony diameter, biomass and density of spores in addition of capability to produce the hydrolytic enzymes for cellulose.
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Microorganisms have an active role in biotechnology for example yeasts, especially in some genus like Saccharomyces, Pichia, and Candida. C.tropicalis one of the most important species of Candida and despite it is one of the causative agents of candidiasis but it has a major role in the production of many chemical compounds. C.tropicalis in the previous study was isolated from sheep dung and morphologically and molecularly classified the result of sequencing was elucidate 100% similarity between the studied isolate and other isolates inserted in DNA Data Bank of Japan DDBJ, physiologically this isolate tolerated 6% ethanol concentration in broth media with the ability to the pro
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This article investigates Iraq wars presentation in literature and media. The first section investigates the case of the returnees from the war and their experience, their trauma and final presentation of that experience. The article also investigates how trauma and fear is depicted to create an optimized image and state of fear that could in turn show Iraqi society as a traumatized society. Critics such as Suzie Grogan believes that the concept of trauma could expand to influence societies rather than one individual after exposure to trauma of being involved in wars and different major conflicts. This is reflected in Iraq as a country that was subjected to six comprehensive conflicts in its recent history, i.e. less than half a century; th
... Show MoreThe present work aims to study the possibility of utilization a forward osmosis desalination process as an alternative method to extract water from brine solution rejected from reverse osmosis process.
Experiments conducted in a laboratory–scale forward osmosis (FO) unit in cross flow flat sheet membrane cell yielded water flux ranging from (0.0315 to 0.56 L/m2 .min) when using CTA membrane,and ranging from (0.419 to 2.785 L/m2 .min) for PA membrane under 0.4 bar. Two possible membrane orientations were tested. Sodium chloride with high concentrations was used as draw solution solute. The effect of membrane orientation on internal concentration polarization (ICP) was studied. Two regimes of ICP; dilutive and concentrative were desc
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Most of the water pollutants with dyes are leftovers from industries, including textiles, wool and others. There are many ways to remove dyes such as sorption, oxidation, coagulation, filtration, and biodegradation, Chlorination, ozonation, chemical precipitation, adsorption, electrochemical processes, membrane approaches, and biological treatment are among the most widely used technologies for removing colors from wastewater. Dyes are divided into two types: natural dyes and synthetic dyes.
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