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.
The mucilage from the seeds of Lallemantia royleana family Labiatae was extracted and subjected to preformulation study for evaluation of its suitability for use as suspending agent. Furosemide suspensions were prepared using (1.5% w/v) of the extracted Lallemantia royleana mucilage, (1.5% w/v) chitosan and (0.35% w/v) xanthan gum. The mucilage was white in color and the average yield of dried mucilage obtained from L.royleana nutlets was 14 % w/w of the seeds used. It is sparingly soluble in water but swells in contact with it, giving a highly viscous solution. It is slightly acidic to neutral. It was found that the extracted natural mucilage of Lallemantia royleana exhibited a higher viscosity profil
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This work was conducted to study the extraction of pelletierine sulphate from Punica granatum L. roots by liquid membrane techniques. Pelletierine sulphate is used widely in medicine. The general behavior of extraction process indicates that pelletierine conversion increased with increasing the number of stages and the discs rotation speed but high rotation speed was not favored because of the increased risk of droplet formation during the operation. The pH of feed and acceptor solution was also important. The results exhibit that the highest pelletierine conversion was obtained when using two stages,(10 rpm) discs speed of stainless steel discs,(pH= 9.5) of feed solution and (pH= 2) of acceptor solution in n-decane. Assuming the existence
... Show MoreThis study is about awareness of teaching explanation difficulties in the Islamic university from the lecturers point of view. It discussed the difficulties and the traditional teaching methods of explanation. The study concentrated on teaching Islamic studies in general and teaching explanation in specific and set difficulties so as to be treated.
The study is of three chapters, the first contains the difficulties in several aspects like the educational goals, text contents, teaching methods and styles, students, educational techniques, educational aids and evaluation, it addition to the lecturers of Islamic university colleges in 2009-2
... Show MoreIn this work, the extraction of glycyrrhizin from Licorice using bulk liquid membrane technique was developed and optimized. The effect of various parameters such as pH of stripping and donor solutions, temperature, stirring speed and kinetic parameters were investigated. Moreover, to study the impact of the polarity of membrane solvent, two types of extraction solvents were used as a membrane solvent: n-Hexane was used as a non-polar solvent and 1-Hexanol was as a polar solvent. The optimum extraction condition was found (95.53%) using 1-Hexanol, rotating speed was 400 rpm, and pH of the acceptor and donor solutions were 8 and 5.5, respectively. The reaction kinetics constants ( and ) for the transport of glycyrrhizin from the donor pha
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In this work, the nano particles of Na-A zeolite were synthesized by sol –gel method. The samples were characterized by X-ray diffraction (XRD), X-ray luorescence (XRF), Surface area and pore volume, Atomic Force Microscope (AFM) and Fourier Transform Infrared Spectroscopy (FTIR). Results show that the nano A zeolite is with average crystal size is 74.77 nm., Si/Al ratio 1.03, BET surface area was 581.211m2/g and the pore volume for NaA was found equal to 0.355cm3/g.
Background: Neonatal septicemia is a significant cause of morbidity and mortality worldwide especially so in developing countries. To reduce the mortality caused by neonatal septicemia, it became vital to diagnose it as soon as possible and treat with administration of appropriate antibiotics.Objective: To study the relationship between themicroorganisms isolated from septicemic neonates with place of delivery.Patients and Methods: Blood sample was obtained from 76 neonates (50 of them are born in Baghdad teaching hospital (Inborn), 26 of the babies are born at home or in Al-Elwya teaching hospital (out born) ,the laboratory diagnosis for the out born patients done in the same hospital(Al-Elwya teaching hospital .The aged of the neo
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This study was carried out to find out the effect of germination of broad beans and chickpeas seeds for different periods on the sensory properties of homus bethina and falafel. The results revealed that the studied properties were significantly different (P<0.05) in tenderness, flavor and overall acceptance as compared to control samples. While other properties such as appearance, body (texture), leavening and color did not showed significant differences.It was found that treatment B1 (100% germinated broad beans) varied significantly in tenderness in comparison with control samples.Treatment B3 (75% ordinary bread beans + 25% germinated broad beans) revealed significant differences (P<0.05) in both flavor and overall acceptance as compar
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In this paper, we have extracted Silica from rice husk ash (RHA) by sodium hydroxide to produce sodium silicate. 3-(chloropropyl)triethoxysilane (CPTES) functionalized with sodium silicate via a sol-gel method in one pot synthesis to prepare RHACCl. Chloro group in compound RHACCl replacement in iodo group to prepere RHACI. The FT-IR clearly showed absorption band of C-I at 580 cm-1. Functionalized silica RHACI has high surface area (410 m2/g) and average pore diameter (3.8 nm) within mesoporous range. X-ray diffraction pattern showed that functionalized silica RHACI has amorphous phase .Thermogravemitric analysis (TGA) showed two decomposition stages and SEM morphology of RHACI showed that the particles have irregu
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