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.
A study of the hadiths of supplication orchestration
Gypseous soil is a collapsible soil, which causes large deformations in buildings that are constructed on it. Various methods have been used to minimise this effect, such as replacing the gypseous soil or using soil stabilisation (grouting or soil improvement). This study was carried out on four types of gypseous soils that have different properties and various gypsum contents. The testing was carried out on remoulded samples to evaluate the compressibility of gypseous soil under different conditions. The samples were grouted with acrylate liquid. The relationships between the injection pressure and the radius of flow, between time of injection and radius of flow, and between time and quantity of acrylate liquid are investigated on
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Drama "The Condamned of Altona" shows many different aspects of the 20th century. It contrasts strong, self-assured characters with weak, unsure ones to address absurdism and existentialism. Also, this piece shows the misery of the decade by showing Frantz, one of the play's heroes, taking his own life. Through suicide, disillusionment, and worry, the play depicts the upheaval of the 20th century, which was marred by numerous crises. The play's depiction of the jail and its inmates exemplifies the regret that accompanied the two world wars . Not only does guilt serve as a metaphor for the wrongs done by the Germans, but it also serves as a metaphor for the French inactivity during the occupation. And also the writers who did not devote thei
... Show MoreThe housing sector in Iraq has suffered from multiple problems, the most prominent of which is the failure of the housing market to achieve housing production that faces population increases and a high number of households.
The research aims to foresight the future of the housing sector, and to identify the expectations of experts in the development of the housing sector and housing production, in order to overcome the obstacles and problems.
Foresight does not mean forecasting about future events, but rather it is a process and skill aimed at familiarity with providing sufficient knowledge to achieve the desired future goals, Accordingly, adopting a method of future foresight will reduce error, surprise and s
... Show MoreStandards of audit have been defined issued them by professional organizations the audit risk is: Failure of the auditor inadvertently to amend his opinion on the financial statements in suitable method, although these statements are Interpolated Essentially. As result the deep impacts caused by electronic operating systems in the accounting data in the audit process which audit risk has gained attention of many professional sides, especially the audit process and quality is relating with level of discovery the auditor for the mistakes of origin (misrepresentations) all their types and give the necessary confidence for the auditor to express his technical opinion in fidelity and certified financial statements which prepared electronicall
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Seven species of semi venomous Opisthoglypha snakes (Reptilia, Ophidia) of Iraq are listed with important characteristics in morphology due to geographical and individual variation of species, as well, the confusion in the scales count of Telescopus tessellatus martini (Schmidt, 1939) of Iraq are discussed.
Biological Activity of Complexes of Some Amino Acid
Back ground: Spectral analyses of solutions have long been applied to various body fluids for the purpose of clinical study as well as research .Human saliva can be easily obtained by non invasive .In this study typical spectra (for UV and IR) of saliva of oral cancer Squamous cell carcinoma patients were determined under average conditions and evaluated in relation to the spectra of normal specimens. Materials and Methods: Seventeen patients of oral cancer Squamous cell carcinoma and seventeen age matched healthy subject were included in this study .Chewing - Stimulated Saliva was collected in plastic test tube and stored at -20° C. Bach of saliva samples were used for UV and IR measurements. Results: Many differences between the IR spect
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