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.
Abstract Background: Coccidiosis in birds, including ostriches, is a major concern for animal health and productivity worldwide. There is limited data on Eimeria infections in farmed ostriches (Struthio camelus). Therefore, this research aims to investigate the species of Eimeria in ostriches using molecular methods. It is considered the first detailed molecular study of Eimeria species in Iraqi ostrich farms. Methods: A total of 150 ostrich feces samples from farms in three regions of Iraq using both microscopy and molecular methods. Statistical analysis of the positive samples was performed using the chi-square test, with the significance level set at a p-value < 0.05. Results: Initial testing showed a high prevalence rate of 51.33%. Five
... Show MoreFluidization process is widely used by a great assortment of industries worldwide and represents a trillion dollar industry [6]. They are currently used in separation, classification, drying and mixing of particles, chemical reactions and regeneration processes; one of these processes is the mass transfer from an immersed surface to a gas fluidized bed
The national educational systems both in the Russian Federation and Iraq Republic have to adjust the training programs to duly prepare the pedagogical university students for the modern challenges and situations on the market of educational services. For success of the service, the education specialists have to fully mobilize their physical, mental and emotional resources and persistently advance their skills and knowledge using the relevant online education courses; practical research conferences; persistent self-education to master theoretical fundamentals of the modern physical education and sport service; and be active in trainings and competitions in their vocational individual game sports including badminton, table tennis and tennis.
... Show MoreBackground: Isoxazoles are an important class of five-membered unsaturated heterocyclic compounds. They show several applications in diverse areas such as pharmaceuticals, agrochemistry and industry. Isoxazoles are also found in natural sources showing insecticidal, plant growth regulation and pigment functions. Current study was conducted for synthesis of twenty five new Isoxazole derivatives and to evaluate the in vitro antibacterial activities of these derivatives. Methods: Benzaldoxime and their substituted [I] ae were prepared via addition-elimination reactions between aromatic aldehyde and hydroxylamine hydrochloride. In a second step, para-or meta-substituted benzaldoximes [I] ae were reacted with N-chlorosucceinimide in DMF to yield
... Show MoreThis study was aimed to investigate the response surface methodology (RSM) to evaluate the effects of various experimental conditions on the removal of levofloxacin (LVX) from the aqueous solution by means of electrocoagulation (EC) technique with stainless steel electrodes. The EC process was achieved successfully with the efficiency of LVX removal of 90%. The results obtained from the regression analysis, showed that the data of experiential are better fitted to the polynomial model of second-order with the predicted correlation coefficient (pred. R2) of 0.723, adjusted correlation coefficient (Adj. R2) of 0.907 and correlation coefficient values (R2) of 0.952. This shows that the predicted models and experimental values are in go
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Due to the importance of the extraction process in many engineering and medical industries, in addition to great interest in medicinal plants, in this research, microwave-assisted extraction has been applied to extract some active compounds from Rosmarinus officinalis leaves. The optimal extraction conditions were then determined by calculating the ratio and extraction efficiency. The process has also been described through kinetic study by applying five kinetic models, the Hyperbolic diffusion model, Power low model, the First order reaction model, Elovich's model, and Fick's second law diffusion model and determining their compatibility with the studies operation, and determining the kinetic constants for each model. The result
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Background: Determination of local bone mineral density (BMD) immediately after implant insertion play an important role in implant success rate, may offer comprehensive description of the bone, and give enough information to the surgeon prior to implant insertion and at follow up status. The aim of the present study is to evaluate the changes of local bone density in the dental implant recipient sites by using computerized tomography. Material and method: The sample consisted of (20) dental implants recipient sites, bone density assessment was done twice, immediately after implants insertion and after six months. Results: The mean HU of the bone around the implant insertion site, immediately after implant placement was 552.28 HU, and inc
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The nuclear density distributions and size radii are calculated for one-proton 8B, two-proton 17Ne, one-neutron 11Be and two-neutron 11Li halo nuclei. The theoretical outlines of calculations assume that the nuclei understudy are composed of two parts: the stable core and the unstable halo. The core part is studied using the radial wave functions of harmonic-oscillator (HO) potentials, while the halo is studied through Woods-Saxon (WS) potential. The long tail behaviour which is the main characteristic of the halo nuclei are well generated in comparison with experimental data. The calculated size radii are in good agreement with experimental values. The elastic electron scattering form factors of the C0 component are also c
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