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 the present research, the chemical washing method has been selected using three chelating agents: citric acid, acetic acid and Ethylene Diamine Tetraacetic Acid (EDTA) to remove 137Cs from two different contaminated soil samples were classified as fine and coarse grained. The factors that affecting removal efficiency such as type of soil, mixing ratio and molarity have been investigated. The results revealed that no correlation relation was found between removal efficiency and the studied factors. The results also showed that conventional chemical washing method was not effective in removing 137Cs and that there are further studies still need to achieve this objective.
The tracking of satellites motion and their path around the earth is important things in the mechanical of satellites motion. Significant parameters for the determination of time entrance and existence of the satellite could be obtained from the shadow of the earth. In the present work the tracking and time determination for entry and exit from earth shadow have been studied. In the present work we built a software for tracking the motion of satellites in orbit around the earth and determine the change of both distance and speed as a function of time. The perturbations effect on the satellite has been neglected from the earth atmosphere drag and the earth gravity and other effects. The equation for calculating the shadow is solved using num
... Show MoreThe extraction of iron from aqueous chloride media in presence of aluminum was studied at different kinds of extractants(cyclohexanone, tributyl phosphate, diethyl ketone), different values of normality (pH of the feed solution), agitation time, agitation speed, operating temperature, phase ratio (O/A), iron concentration in the feed, and extractant concentration]. The stripping of iron from organic solutions was also studied at different values of normality (pH of the strip solution) and phase ratio (A/O). Atomic absorption spectrophotometer was used to measure the concentration of iron and aluminum in the aqueous phase throughout the experiments.The best values of extraction coefficient and stripping coefficient are obtained under the
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A new series of Sulfamethoxazole derivatives was prepared and examined for antifibrinolytic and antimicrobial activities. Sulfamethoxazole derivatives bear heterocyclic moieties such as 1,3,4-thiadiazine {3}, pyrazolidine-3,5-diol {4} 6-hydroxy-1,3,4-thiadiazinane-2-thione {5} and [(3-methyl-5-oxo-4,5-dihydro-1H-pyrazol-4-yl)diazenyl] {8}. Their structures were elucidated by spectral methods (FT-IR, H1-NMR). Physical properties are also determined for all compound derivatives. Recently prepared compounds were tested for their antimicrobial activity in the laboratory. Each screened compound showed good tendency to moderate antimicrobial activity.
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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
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Background: Cholera has been recognized as a killer disease since earliest time. The disease is caused by infection of the small intestine by Vibrio cholerae O1 and O1391 which is characterized by severe dehydrating diarrheal condition and is one disease in modern times that is epidemic, endemic and pandemic in nature. Objective: This study was carried out to detect and isolate V. cholerae from patients suffered from watery diarrhea, which may cause severe complications such as dehydration, shock followed by death. Materials and methods: stool specimens were collected from 308 patients with watery diarrhea. These samples were tested with many criteria such as TCBS agar, gram stain, biochemical tests and VITEK-2 system to improve the isolati
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From different hospitals in Baghdad city, 25 clinical isolates of Proteus spp. were collected from different clinical samples, all isolates were identified as Proteus mirabilis by using bacteriological and biochemical assays in addition to Vitek-2 identification system. 15 (60%) isolates were identifying as Proteus mirabilis. The susceptibility of P. mirabilis isolates towards cefotaxime and ceftazidime was (66.6 %), (20%) consecutively; while extended spectrum β-lactamases producing P. mirabilis percentage was (30.7 %). Because blaVEB-1 was documented as an important indicator for increasing risk of extended spectrum beta ßlactamases producing P. mirabilis isolates that began to spread from many geographic area to Far east which inc
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