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.
Background Cadmium (Cd), one of the most abundant heavy metals, is extremely toxic to both humans and animals. hIt is well known that zinc (Zn) administration reduces Cd-induced toxicity and that metallothioneins can have a protective effect in biological systems to mitigate Cd toxicity. Objective The aim of the current study to determine if Zn administration affected the induction of MT-1 and MT-2 in the liver tissue in mice exposed to Cd. Materials and methods Metallothionein protein (MT) level in the tissue of male mice were detected using the anion -exchang high-performance liquid chromatography coupled (HPLC)assay and immunohistochemical staining. Results Single treatment to zinc or cadmium increase the level of MT in the liver, but zi
... Show MoreThe purpose of this study was to know the effect of adding different percentages of soy protein a humidifier to mix Sarcoma Barker component of the beef mixture Alkhalt added Alber elderly chicken with five ratios were manufacturing Alberkr discs for sensory evaluation
The objective of this study is to determination the content of some heavy metals (lead, cadmium, chromium) in colored tattoo stickers. twelve kinds of colored tattoo stikcers were collected from Baghdad markets, it was estimated heavy metals using atomic absorption spectrophotometer (Shimadzu A5000). The results indicated the concentrations of lead in all samples (1.61_1.00 mg / kg) and chromium in the three samples (0.85_0.97 mg / kg) while other samples are free of chromium , and cadmium. These elements are the components of printing inks and dyes in tattoo stickers, and this does not conform to the health and safety conditions for the packaging of food according to the organizations of the health and safety of
... Show MoreThe aim of this study was to increasing natural carotenoides production by a locally isolate Rodotorula mucilagenosa M. by determination of the optimal conditions for growth and production of this agents, for encouragest to use it in food application permute artificial pigments which harmfull for consumer health and envieronmental. The optimal condition of carotenoides production from Rhodotorula mucilaginosa M were studied. The results shows the best carbon and nitrogen source were glucose and yeast extract. The carotenoids a mount production was 47430 microgram ̸ litter and 47460 microgram ̸ litter, respectively, and the optimum temperature was 30°C, PH 6, that the carotenoides a mount was 47470 microgram ̸ litter and 47670 microgr
... Show MoreThis study was conducted to evaluate the bottled water quality for the six-producing companies in Baghdad city, where selected six brands which are the most marketed in the Iraqi market, especially in Baghdad, where taking the proper amount of bottled water in September 2015 and included the studied characteristics (EC , pH ,TDS, Turbidity, Ca+2, Mg+2, Cl-, No3-, So4-2, HCO3-, Na+ and K+) in addition to the total population of bacteria aerobic and coliform, and compare the results with the standard specifications of the Iraqi and the World Health Organization (WHO), as well as to compare the results of sampling specifications mentioned on the packaging by the producing companies. The results showed the presence of high significant differ
... Show MoreBackground. Bone healing is a complex and dynamic process that represents a well-orchestrated series of biological events of cellular recruitment, proliferation, and differentiation. The use of medicinal plants in bone healing has attracted increasing interest because of their lower side effects. Punica granatum seed oil (PSO) contains high levels of phenolic compounds, promotes osteoblast function, and plays an important role in bone remodeling. A gelatin sponge (Spongostan) is a hemostatic agent that is extensively applied as scaffolds in engineering and as drug carriers in the medical field. This study aimed to evaluate the effectiveness of PSO for bone healing enhancement. Twenty adult male New Zealand rabbits, weighing an avera
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Make a search on the vegetative parts of the plant local horse guilt of some elements in the Haj Omran area in northern Iraq has included recognition of certain nutrients
60 patients diagnosed as having urticaria were included in the study ; 30 patients were effected with acute urticaria and 30 patients were affected with chronic urticaria. In addition, 30 healthy adult volunteers were selected as control group .The patients and control groups sera were examined with enzyme linked immunosorbent assay ( ELISA) to detect total level IgE and radial immunodiffusion (RID) to detect levels of IgG , IgA and IgM . The total level of IgE in acute urticaria ( 1.45±0.13) IU/mL and chronic urticaria (2.12 ± 0.10) IU/mL patients were significantly higher than the control groups ( 0.85 ± 0.10)IU/mL (p<0.05). The level of IgG in acute urticaria ( 12.5± 0.42) g/L and chronic (13.16±0.40) g/L patients , IgA in acute (2.
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Industrial 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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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
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