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, synthesised new ligand: potassium 2,2'-(quinoxaline-2,3- diyl)bis(1-phenylhydrazinecarbodithioate) (L). The ligand synthesised by reacting N1,N2-dip-tolyloxalamide as the starting material with CS2 and KOH to add the CS2 group and then with phenylendiammine to achieve (L). The ligand used in the synthesis of complexes with (CoII, NiII and CdII). The new ligand and its complexes characterised by FT-IR, UV-Vis, 1H, 13C-NMR, Mass spectroscopy, and elemental analysis, in addition to the above techniques were using magnetic moment, atomic absorption, chloride content, and melting point to describe the metal complexes.
This study was designed for isolation and molecular identification of Nontuberculous Mycobacterium (NTM) from fish during the period between October and December 2017 from Karbla province, Iraq. This study included 200 fresh fish samples from four different species including Spondyliosoma cantharus, Liza abu, Carassius carassius and Cyprinuscarpio. Three samples of each fish were taken including gills, muscles and all internal organs. The samples were processed by decontamination, concentration of 4% sodium hydroxide, and 0.1 ml of sediment was streaking on Löwenstein Johnson (LJ) media; then the bacterial cultures were incubated at 28-30 °C for 3days up to 4 weeks and suspected colonies were stained with acid fast stain to confir
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The fresh water green alga Chlorella vulgaris was selected to study its ability to degrade one of polycyclic aromatic hydrocarbon (PAHs)known is anthracene (ANT). ANT is widely used in artificial products such as wood preservatives; insecticides; dye and coating materials. This algae was cultivated in CH-10 medium under constant laboratory conditions and exposed to different concentrations (1, 3, and 5 mg/l) of anthracene for 3, 5, 7, 9 and 15 days, with the concentration of ANT measured by high performance liquid chromatographic analysis (HPLC). The results showed that C.vulgaris has high ability to reduce anthracene to 80% at 1mg/Lcon centration after 3 days and 100% after 5 days, while at 3 and 5 mg / L concent
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The present study provides the first record species of the genus Lithobius Leach, 1814, L. ferganensis (Trotzina, 1894) which was collected from the middle of Iraq. A detailed explanation of the morphology and the diagnostic characters of specimens of both sexes is provided.
This present work adopts the analytic and descriptive methodology to recognize and analyze the major challenges faced by the Iraqi government. The study focuses on the press and national partiers’ cabipility of changing and unifying the opinions of the public in a way that serves the Iraqi interest, taking into account the fact that the press is the fourth authority after the three major authorities, (Executive, Legislation and Judicial). The researcher has concluded that the Iraqi press and parties initiated a national front with the Iraqi people against the government’s signature of the treaty of (1930). The national front illustrated that signing the treaty strengthens the authority of the British occupation u
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Celery and coriander are vastly applied in modern medicine and traditionally because various medicinal and nutritional benefits depend on their medicinal characteristics. The study aimed to detect, isolate and compare extracts contents of phenolic acids (caffeic and p-coumaric acids) in ethyl acetate fraction of fresh and dry aerial parts of coriander (Coriandrum sativum L.) and celery (Apium graveolens L.) of the Apiaceae family. The extraction of these constituents was carried out by maceration method using 70% ethanol and fractionation was done by using petroleum ether, and ethyl acetate. The existence of caffeic and p-coumaric acids in aerial part extracts of two plants was identified by thin-layer chromatography (TLC) and high-
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The kinetic of atropine pertraction from seeds of Datura Metel Linn plant was studied. Diisopropyl ether, n-hexane and n-heptane were used as membranes for atropine recovery. The effect of speed of agitation and time in the range of 200-300 rpm and 0-3.5h, respectively were studied using the proposed membranes. The pertraction experiments were carried outs in a batch laboratory unit. The liquid-liquid pertraction was found to be very suitable for atropine recovery from its liquid extracts of Datura Metel seeds. A high purity (94-96%) can be obtained in the receiver phase. The pertraction process was found to be very selective for atropine recovery with diisopropyl ether membrane. As the speed of agitation increases the efficiency of pertrac
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The present study was conducted to determine the optimum conditions required for lipase enzyme activity extracted from germinated sunflower seeds, including temperature, pH, agitation, time of incubation, enzyme concentration, substrate type, and concentrations of mineral salts and EDTA. Optimum pH, temperature and time of incubation required for lipase stability were also determined. The results showede optimum lipase activity (3.251U/ml) wasund at 30 ÌŠC and pH 7 after 20 minutes of incubation when using 1 ml lipase enzyme with 0.02 ml of CaCl2 (10 mM) at 100 rpm of agitation and in the presence of olive oil as the substrate for enzyme reaction. EDTA appeared to have inhibitory effects, while Ca+2 and Mg+2 have stimulatory effec
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In the present work, it had been measured the concentration of radon gas (CRn) for (10) samples of cement used in constructions before and after painting them using enamel paint, purchased from the local markets, to see the extent of its ability to reduce emissions of Rn-222 in the air. These samples were obtained from different sources available in the local markets in Baghdad and other provinces. The measurements were done by the American-made detector (RAD7). The results showed that the highest CRn in the air emitted from cement samples after coating was in the cement sample (Iranian origin) where the concentration was (58.27 Bq/m3) while the lowest CRn was found in building material samples
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