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
... Show More
(11)
(9)
The study included the determination of pollen grains features for 8 genera and 13 taxa of Mimosoideae subfamily grown in Baghdad/ Iraq by using each of light and scanning electron microscope. The samples of taxa were collected from various sites in Baghdad province in central Iraq located on 32 45° 0-33 45 0 N and 44 0 0- 44° 45 0 E. the results from this study revealed different pollen types as monad in each of Leucaena, Prosopis, and Neltuma, tetrad in Mimosa and polyads in Acacia, Albizia, Calliandra, Pithecellobium and Vachellia. Each taxa of these genera characterized by special palynological features as shape, size, number of polyads grain and conplateuration as well as other parameters included other dimensions, and these
... Show More
Samples of the green algae were collected from water of Shatt al-Arab in Garmat Ali in Basra. After purification, the green algae identified on Enteromorpha sp. The samples were dried and milled, then sulfated polysaccharides were extracted with hot water at 90°C precipitated with absolute ethanol, dialysed and lyophilized. The chemical composition was total sugars 56.4%, protein 1.3% and sulfur 19.7%. Antioxidation activity of sulfated polysaccharides was studied by four method and included estimation of ability of scavenging hydroxylated radicals, the results showed an increased in ability with increasing concentrations. Ability of scavenging and was 59.86% at the concentration of 2.5 mg/ ml, but BHT was 81.36%. Ability of scavenging
... Show MoreThe 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 More
(13)
(10)
Hybrid bilayer heterojunction Zinc Phthalocyanine (ZnPc) thin-film P-type is considered as a donor active layer as well as the Zinc Oxide (ZnO) thin film n-type is considered as an acceptor with (Electron Transport Layer). In this study, using the technique of Q-switching Nd-YAG Pulsed Laser Deposition (PLD) under vacuum condition 10-3 torr on two ITO (Indium Tin Oxide) and (AL) electrodes and aluminum, is used to construct the hydride bilayer photovoltaic solar cell heterojunction (PVSC). The electrical properties of hybrid heterojunction Al/ZnPc/ZnO/ITO thin film are studied. The results show that the voltage of open circuit (V_oc=0.567V), a short circuit (I_sc=36 ?A), and the fill factor (FF) of 0.443. In addition, the conversion
... Show More
Background: Diabetes mellitus is one of the commonest chronic disorders worldwide with a rapid rise in prevalence. In Iraq its prevalence is high especially in elderly age group. Patients with type 2 diabetes mellitus have higher vulnerability for complications, whether microvascular or macrovascular. Ocular complications are common in diabetes mellitus, and comprise diabetic retinopathy, diabetic papillopathy, cataract, glaucoma, dry eye disease and diabetic keratopathy. Diabetic keratopathy involves endothelial and epithelial tissues of the cornea, leading to persistent epithelial defect, corneal erosion, or corneal ulcers.
Aim of the Study: To compare the mean corneal endothelial cell count between patients wi
... Show More
(1)
(15)
The present study is concerned with studying the effect of aquatic plant Hydrella vorticellata with the concentration of 10 and 20 gm/2 K gm soil on percentage and growth rate of germinating seeds of Hordeum vulgare and Vicia faba. More overs, the quantitative amount of NPK in both tested plants and Hydrella vorticellata, are estimated as an organic fertilizer. It has also been find the total number of root cells, the number of dividing cells, and stages of mitosis. The study reveales, that there are no significant differences between the concentration of hydrella used in germination percentage, growth rate, wet and dry weight. While there are differences in the plants containing NPK. The number of cells dividing stages and number of divid
... Show More
Staphylococcus Sp.is the most common type of bacteria found in contamination place, we design this
study to compare the contamination accident between two hospitals in Baghdad.One of them isthe Burns
Specialist Hospital in the Medical CityinRusafa and another one is Al-Karama Hospital in Karkh. The
samples were collected fromOperativeWard No1 (OW1), Operative Ward No2 (OW2), Consulting Pharmacy
(CP), Emergency Room (ER), Reception Room (RR), Women's Ward (WW) and Men's Ward (MW).The
samples were taken from inside each clinical unit, surfaces, food, and air. The results showed that the
number of samples containing Staphylococcus sp. bacteria is 81, including 45 belonging to Al-Karama Burns
Ward Ho
(1)