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.
This study aimed to investigate the feasibility of treatment actual potato chips processing wastewater in a continuously operated dual chambers microbial fuel cell (MFC) inoculated with anaerobic sludge. The results demonstrated significant removal of COD and suspended solids of more than 99% associated with relatively high generation of current and power densities of 612.5 mW/m3 and 1750 mA/m3, respectively at 100 Ω external resistance.
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The study aimed to assess Milk Thistle oi reducing the aflatoxin damage and improve the health status of local breed hens. The study was performed on 60 hens aged 35 weeks, weighing 2-2.5 kg of the body was obtained and housed on a private poultry farm in north of Baghdad (Altaji) for a period of 9 weeks from 22 October until 22 December 2020. Hens were divided to three treatments (20 hens). The first was fed a basal diet free of aflatoxin. The 2nd fed on a contaminated diet with aflatoxin (14.6 ppb) while the 3rd fed as in the 2nd with 0.5% of Milk Thistle oil /kg feed. A 5 hens from each group were slaughtered at the end of the trial for the histopathological inspection of liver, spleen and intestine tissues to evaluate the harmfu
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Rutting is one of the major distresses in pavement. The objective of this paper is to develop an improved asphalt binder grading system for Iraq based on the principal of Superpave system, and increasing performance grade of product asphalt binder in Iraq using polymers without raising the viscosity of the binder. Two types of polymers are used, Plastomers, Functionalized Polyethylene (PE) which is developed by asphalt research group in Wisconsin University in the USA, and Elastomers, Styrene Butadiene Styrene (SBS) with and without cross linker. Mastercurve are drown for these modified binders, Rolling thin film aged, to show effects on rheological properties at high temperature for complex modulus (G*) and phas
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Human resources are considered as strategic fortune for being the main driver of the development wheel in the society, and the field of education and learning is one of the main pillars of this fortune for its great effect in the process of economic and social progress of individuals. I was the subject of education to the concerns of many countries, as adopted national policies . And regional support and the reduction of constraints, so came our study (education hub for human development) to identify the role of education in human development and to identify the obstacles facing the education process and the extent of its impacts negatively on the process of human development also contribute to the knowledge of school enrollment and the
... Show MoreIndustrial effluents loaded with heavy metals are a cause of hazards to the humans and other forms of life. Conventional approaches, such as electroplating, ion exchange, and membrane processes, are used for removal of copper, cadmium, and lead and are often cost prohibitive with low efficiency at low metal ion concentration. Biosorption can be considered as an option which has been proven as more efficient and economical for removing the mentioned metal ions. Biosorbents used are fungi, yeasts, oil palm shells, coir pith carbon, peanut husks, and olive pulp. Recently, low cost and natural products have also been researched as biosorbent. This paper presents an attempt of the potential use of Iraqi date pits and Al-Khriet (i.e. substances l
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Background: Monocyte chemotactic protein-1 (MCP-1) is a chemokine expressed by inflammatory and endothelial cells. It has a crucial role in initiating, regulating, and mobilizing monocytes to active sites of periodontal inflammation. Its expression is also elevated in response to pro-inflammatory stimuli and tissue injury, both of which are linked to atherosclerotic lesions. Aim of the study: To determine the serum level of MCP-1 in patients with periodontitis and atherosclerotic cardiovascular disease in comparison to healthy control and evaluate the biomarker's correlations with periodontal parameters. methods: This study enrolled 88 subjects, both males and females, ranging in age from 36-66 years old, and divided into four groups: 1<
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Background: Orthodontic force is considered to stimulate cells in the periodontium to release many mediators such as cytokines which play a responsible role for periodontal and alveolar bone remodeling, bone resorption and new bone deposition. Aim of this study was carried out to estimate changes of the (interleukin-one beta, tumor necrosis factor – alpha and C-reactive protein) levels in unstimulated whole saliva during the leveling stage of orthodontic tooth movement. Materials and methods: The sample consisted of thirty adult patients (12 males and 18 females) with ages ranges (19-23) years. Each sample had Class I and Class II malocclusion dental classification and required bilateral extraction of their maxillary first premolars, und
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The study showed significant differences between the average weight lens and the average amount protein in the lens between that Kestrel Falco tinnunculus L. and the Collared Dove Streptopelia decaocto F. , also the study electrical migration of lens proteins having one bundle of crystalline –? in Kestrel compared with three bundles in Collared Dove, two bundles of crystalline – ? in both , and crystalline – ? appeared as one bundle in both birds.
The absence of ecological perception in the local urbanization resulted in the lack of a clear conception of achieving sustainability in its simplest form in the urban reality and in the city of Baghdad in particular. The research assumes the possibility of achieving urban sustainability in Iraqi cities by applying the cities for the most effective methods to implemented ecological solutions and introducing appropriate urban planning tools and improve the living environment. The research focuses on the ability to define some aspects to achieve a sustainable local urban identity from global experiences. This was performed by proposing a scheduled theoretical framework, through which the features of sustainability can be extrapolated from the
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