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 presence of residual antibiotics in water results in the development of antibiotics resistant genes. The available wastewater treatment systems are not capable of removing such antibiotics from sewage. Thus, antibiotics need to be removed before the discharge of wastewater. Adsorption is among the promising techniques for the wastewater treatment to aid the removal of a wide range of organic and inorganic pollutants. The present work is a contribution to the search for an economical method for the removal of low concentrations of amoxicillin (AMX) from water by adsorption on water treatment residue, WTR, taken from a local drinking water facility. The chemical composition and the adsorptive characteristics of the material were first
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In this study miconazole nitrate was formulated as topically applied emulgel; different formulas were prepared using sodium carboxymethylcellulose (SCMC) and carboxypolymethylene (carbomer 941) as gelling agents. The influence of type of gelling agent and concentration of both oil phase and emulsifying agent on drug release was studied and compared with commercially available miconazole nitrate cream (Mecozalen®). The results of in vitro release showed that SCMC emulgel bases gave better release than carbomer 941 bases and the release of drug increase from both bases as a function of increasing the concentration of emulisifying agent. The oil phase had retardation effect when
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New Schiff bases derived from D-galactose were synthesized by condensation of aldehyde (1,2:3,4-Di-O-isopropylidene-6-carboxaldehyde-α-D-galactopyranose) with different aromatic amines such as (4-bromo, 3-hydroxy, 4-iodo, 4-methoxy) aniline in dry benzene using glacial acetic acid as a catalyst. These compounds were converted to oxazepine derivatives by addition reaction with maleic anhydride in dry benzene as a solvent. The structures of the synthesized compounds have been characterized by elemental analysis, FTIR spectra, some of them by using 1HNMR spectra and measurement of its physical properties.
There are numerous ways to define the traits of influential professors, including behavior (such as warmth, civility, and clarity), knowledge (of subject matter and students), and beliefs, among many others. According to research; influential teaching can also defined as the capacity to improve student achievement. This; is only one way to express effectiveness, as stated. However; the preferred definition of high-quality teaching in the United States and many other countries is teacher effects on student achievement. Therefore; this study aimed to determine the characteristics of influential professors in Online Teaching at the English department. This study; believed to be significant for college professors who will show them how
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Numerical simulations have been investigated to study the external free convective heat transfer from a vertically rectangular interrupted fin arrays. The continuity, Naver-Stockes and energy equations have been solved for steady-state, incompressible, two dimensional, laminar with Boussiuesq approximation by Fluent 15 software. The performance of interrupted fins was evaluated to gain the optimum ratio of interrupted length to fin length (
This study focused on treating wastewater to remove phosphorus by adsorption onto naturaland local materials. Burned kaolin, porcelinite, bauxite and limestone were selected to be testedas adsorption materials.The adsorption isotherms were evaluated by batch experiments, studyingthe effects of pH, temperature and initial phosphorus concentration. The results showed that at pH6, temperature 20°C and 300 mg/l initial phosphorus concentration; the sorption capacity was0.61, 9, 10 and 13 mg/g at 10 h contact time, for burned kaolin, porcelanite, limestone and bauxiterespectively. As the pH increased from 2 to 10 the removal efficiency for the materials differs inbehaviour. The removal efficiency increased from 40 to 90 % for limestone, and dec
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Pharmaceuticals are widely distributed in different applications and also released into the environment. Adsorption of Ciprofloxacin HCl (CIPH) on Porcelinaite was studied at ambient conditions. The adsorption isotherms can be well described using the Freundlich and Temkin equations. The pH of the solution influences significantly the adsorption capacity of Porcelinaite, the adsorption of CIPH increased from the initial pH 1.3 and then decreased over the pH rang of 3.8-9. The adsorption is sensitive to the change in ionic Strength, which indicate that electrostatic attraction is a significant mechanism for sorption process. The enthalpy change (∆H) for the adsorption of CIPH onto Porcelinaite signifies an endothermic adsorption. The ∆G
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