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.
Thermal decomposition of used tires was studied under atmospheric pressure and thermal heating program up to 350 °C for 110-120 min in a glass reactor. The effects of using MgO or SiO2 as catalyst in the thermal pyrolysis of tires waste on the yield ratio, reaction time, initial degree of decomposition also the contents of BTEX compounds in resulting pyrolytic oil was estimated via GC-FID. The results showed that the MgO catalyst gave a gas output in proportion of 17.063%, a liquid in a proportion of 38.245% and a solid product in proportion of 44.733%, while the SiO2 catalyst gave proportions of 14.308%, 40.161% and 45.448% for gas, liquid and solid products respectively. The results revealed that the thermal pyrolysis produced minor cont
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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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Background: Periodontal diseases are inflammatory disorders caused by the accumulation of oral biofilm and the host response to this accumulation which characterized by exaggerated leukocytes and neutrophils attraction to the sites of inflammation by chemoattractants which are a very important part of the pathogenesis of periodontal diseases. This study aimed to determine and compare the clinical periodontal parameters and the leukocyte cell types in the peripheral blood between patients with gingivitis and periodontitis with different severities compared to healthy controls. Materials and methods: This study included 150 male subjects aged between 35-50 years. They were divided into three groups: gingivitis group (n=30), periodontitis p
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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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Measuring the level of communicative competence in news headlines and the level of stylistic and semantic processing in its formulation requires creating a quantitative scale based on the bases on building the scales and their standards. As judging by scientific of journalism studies lies in the possibility of quantifying the journalistic knowledge, i.e. the ability of this knowledge to shift from qualitative language to its equivalent in the language of numbers.
News headlines and editorial processing are one of the journalistic knowledges that should be studied, analyzed stylistically and semantically; their conclusions drawn and expressed in numbers. Press knowledge is divided into two types:<
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The present study introduces description of a new species of genus Arboridia Zakhvaticin 1946, based on a large collection of Cicadellids. External morphological characters particularly male genitalia were discussed and illustrated. The genus Arboridia Zalchvatkiia (Typhlocybinae: Erythroneurini) contains small slender, fragil and attractively coloured and patterned leafhoppers. It was erected by Zakhvatkin in 1946 (Zalchvatkin, 1946). The overall length of adults ranges from 2.5 to 3.4 mm. Members of this genus can be recognized by inner apical cell of forewing which is long with oblique base; Cu confluent with this base at a point near the middle of the length of inner apical cell; two prominent circular deep brown spots on vertex (Zal
... Show MoreThe lesser grain boner Rhizopertha dominica is an important insect on grain in Iraq . in this study phenol and coumarin were isolated from Albizzia lebbek . the addition of there compounds to artificial diet at concentration of 3.4,5% couse high effects on the development of the studed insect larva.the larva were unable to complete its development to adult stage. More over, using 5% coumarin larva were died.Seed germination was not affected when seed were sprayed with concentration of 3,4,5% of coumarin and phenol.
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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