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.
Crude soybean peroxidase (SBP), isolated from soybean seed coats (hulls) at unusually low concentrations, catalyses the oxidative polymerisation of hazardous aqueous benzidine and its 3,3′-dichloro, 3,3′-dimethyl and 3,3′-dimethoxy derivatives in the presence of hydrogen peroxide. The optimum operating conditions for oxidation of 0·10 mM benzidine were investigated. At pH 5, the hydrogen peroxide-to-substrate concentration ratio was 1·5 and the minimum SBP concentration required to achieve at least 95% conversion of the benzidine in synthetic wastewater was 0·43 mU/ml. Progress curves were established for the conversion of the four substrates, and apparent first-order rate constants were derived. Enzyme-catalysed polym
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The purpose of this research is to explain the effect of the dropping of three zeroes from Iraqi dinar on the basic role of the dinar and derivative and dynamic in transaction of Iraqi economic. the importance of this research comes from the effect of this operation on the evaluation of the exchange between the individuals of Iraqi society and the other between the Iraqi individuals and the foreign whom use the Iraqi dinar as a tool for exchange also the effect of the operation on the reserves of the Iraqi dinar the results of this research are as the following:-
1-the operation of dropping of zeroes must be associated with increase of the sharing of economic sectors wi
... Show MoreUsed vegetable oil was introduced to transesterfication reaction to produce Biodiesel fuel suitable for diesel engines. Method of production was consisted of filtration, transesterfication, separation and washing. Transesterfication was studied extensively with different operating conditions, temperature range (35-80oC), catalyst concentration (0.5-2 wt. % based on oil), mixing time (30-120 min.) with constant oil/methanol weight ratio 5:1 and mixing speed 1300 rpm. The concentration of Fatty acid methyl esters (Biodiesel) was determined for the transesterficated oil samples, besides of some important physical properties such as specific gravity, viscosity, pour point and flash point. The behavior of methyl esters production and the phys
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The optimum conditions for production of fibrinolytic protease from an edible mushroom Pleurotus ostreatus grown on the solid medium , Sus medium, composed of Sus wastes (produced from extracted medicinal plant Glycyrrhiza glabra) were determined. Addition of 5% of Soya bean seeds meal in Sus medium recorded a maximum fibrinolytic protease activity resulting in 7.7 units / ml. The optimum moisture content of Sus medium supplemented with 5% Soya bean seeds meal was 60% resulting in 7.2 units / ml.Pleurotus ostreatus produced a maximum fibrinolytic protease activity when the spawn rate,pH of medium and incubation temperature were 2,6 and 30°C, respectively. The maximum fibrinolytic protease activity was 7.6 units / ml when incubat
... Show MoreThis study concluded detection of Toxoplasma gondii in milk, immunologically by using Elisa and nested PCR)nPCR (based on B1 gene, also to investigate the effect of toxoplasmosis, parity, breed and flock on some milk composition in the Iraqi local and Shami goats in the middle of Iraq. A total of 80 milk samples of the lactating goats were collected. Results of this study showed the prevalence of Toxoplasmosis was 21.25% and 28.75% by Elisa and nPCR respectively without significant differences. The sensitivity of Elisa was a low (30.43%) whereas the specificity was a high (82.45%). The degree of agreement estimated by Kappa coefficient revealed a slight agreement (0.14) between two methods. The results indicated that goats infected
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Background: Depression is a state of low mood and aversion to activity, it can affect a person's thoughts, behavior and sense of well-being. It can affect oral health and lead to an increased risk of dental caries. Dental caries is the most common oral infectious diseases that stresses the immune system and causes changes in cellular and molecular components of peripheral blood and C-Reactive Protein is one of these components, considered a key biomarker of inflammation. This study was conducted to assess the effect of depression status on dental caries among 17 years old secondary school female students in relation to salivary C-Reactive Protein. Materials and Methods: A cross sectional study was carried and the whole sample composed of
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The aim of the research is to show the extent of the impact of administrative coordination on municipal performance after the state’s tendency to implement administrative decentralization and transfer of powers (administrative, legal, financial, and technical) from the Federal Ministry (construction, housing, municipalities, and public works) to local governments (governorates), to meet local needs, And since the municipality is considered one of the local administrative institutions and most of its goal is to provide increased municipal services to citizens due to population growth and urban expansion of cities by coordinating their actions using their powers, and in order to address gaps i
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Two local fish Himri Carasobarbus luteus (Heckel, 1843) and Hishni Liza abu (Heckel, 1843) were stained with Alizarin Red and featured some anatomical qualities which cleared the difference of the muscular and skeletal fabric for each fish. Since clear Histologic differences appeared in these two species, it was intended from this study the possibility of adopting a diagnosis between local fish species by staining bones and tissues.