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.
Study Purpose. highlighting the origins of the sport's philosophy. Studying the development of the sport of kendo across different historical periods and its adaptation to societal and cultural changes. Identifying the establishment of the most prominent organizations and federations that contributed to its development, and exploring the sport's basic tools and rules and its most prominent championships. Materials and Methods. The historical approach was adopted in studying the historical process of the development of kendo, as it is appropriate to the nature of the problem. The information has been simplified and presented in a sequential manner that presents the development of the sport of kendo, starting from the origins of its p
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Keys for 22 species representing ten genera Thripidae collection carried out during 1999-2001 in different localities in the middle of Iraq. Of them four species are described as new to science, Frankliniella megacephala sp. nov; Retithrips bagdadensis sp. nov; Chirothrips imperatus sp. nov; Taeniothrips tigridis sp. nov; Another thirteen species are recorded for the first time in Iraq; Thrips meridionalis (Pri.); Microcephalothrips abdominils (Crawford); Scolothrips pallidus (Beach); Scritothrips mangiferae Pri.; Frankliniella tritici Bagnall; Frankliniella schultzie Trybom; Frankliniella unicolor Morgan; Retithrips aegypticus Mar
... Show MoreIn this work, chemical and thermal treatment were used to enhance silica extract on the purity of rice husk and to reduce the impurities associated with the extraction of silica. The thermal degradation of rice husk was studied. The characteristics and thermal degradation behavior of rice husk which investigated using thermogravimetric analyzer (TGA). Hydrochloric acid was used to soak the rice husk and the study of leaching influence is followed by XRF tests for samples before and after the combustion process. Acid treatment and combustion method seem to have a clear effect on silica purity. The pyrolysis processes were carried out at Laboratory temperature up to 650 oC in the presence of nitrogen gas flowing at 150 ml/min. The effect o
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The Diyala River is considered the third most important river in Iraq. However, in the recent period, Diyala Governorate has been subject to several floods. This study aims to simulate an efficient labyrinth weir at the flood escape entrance branching from the Diyala River to reach the best entrance through which the flood waves can pass safely. The discharge coefficient was calculated laboratory for five types of trapezoidal side labyrinth weirs with different sidewall angles. Results showed that the coefficient discharge for the trapezoidal labyrinth side weir with an angle of the sidewall is 75ᵒ and has a discharge coefficient greater than the rest of the labyrinth side weirs. The second part of this study is valida
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Information on the scorpions' fauna of Iraq is limited especially in Thi Qar Province. The scorpion specimens of the present study were collected from the desert area which is located between the provinces of Thi Qar and Al-Muthana (Al-Kata'a region). The Scorpio kruglovi (Birula, 1910) redescribed in this study was found in this area. The diagnostic characters are given and the important features are figured.
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The current research aims to identify: 1) the challenges facing blended education from the point of view of teachers of students with disabilities. 2) The challenges facing blended education from the point of view of teachers of students with disabilities according to the gender variable (males-females). 3) The challenges facing blended education from the point of view of teachers of students with disabilities, according to the academic qualifications of graduates
(institute-bachelors-masters). 4) The challenges facing blended education from the point of view of male and female teachers, according to the functional service period with students with disabilities (less than 8 years - from 9 to 15 years - 16 years and above). 5) the
... Show MoreAll the prepared metal complexes of Pt (IV), Au(III), Rh (III), Co (II) and V(IV) with new ligand sodium [5-(p-nitro phenyl)-/4-phenyl-1,2,4-triazole-3-dithiocarbamato hydrazide] (TRZ.DTC) have been synthesized and characterized in solid state by using flame atomic absorption, elemental analysis C.H.N.S, FT-IR ,UV-Vis Spectroscopy, conductivity and magnetic susceptibility measurements. The nature of the complexes formed in ethanolic solution has been studied following the molar ratio method also was studied stability constant and found to be stable in molar ratio1:1 of VL (IV) and CoL(II) while Pt(IV), Au(III) and Rh(III) complexes stable in molar ratio 1:2 as well as the molar absorptivity for these complexes were calculated. From the prev
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The research problem lies in identifying the challenges facing spreading the culture of peace in Iraq, and the most important factors of its instability from the point of view of leaders and intellectual elites.The research derives its importance from the fact that the peace imposed by justice leads to a stable Iraq and a prosperous future.
The study aims to identify the most prominent challenges and obstacles created by wars and faced the process of spreading the culture of peace and instability in Iraq, and to shed light on the reasons leading to the employment of the capabilities of community leaders in spreading the culture of peace.The objective of the current research is to identify the challenges of spreading the culture o
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The current research aims to identify the problems and needs for both college of political science and college of engineering’s students. The sample was (100) male and female student. The results showed bunch of problems which could be organized descendingly, the scientific domain ranged between (2 - 2.42), the mean of the psychological domain was (2.85), the moral domain ranged between (2.2 – 2.28)m the problems of study earned (2.30), the material domain got (1.95), the medical and social domain obtained (1.925), and finally, the family domain received (1.887).
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Background: Repeated blood transfusion is the main therapeutic option for transfusion-dependent anaemias with consequent iron overload and organ damage .Therefore iron chelating agents are important protective measures for these patients. The aim of this study was to investigate the efficiency and safety of Desferroxamine in paediatrics population subjected to iron overload as a consequence of repeated transfusion in a group of Sudanese children Subjects & Methods: This was a descriptive cross-sectional hospital based study. Conducted in two main paediatric reference hospitals in, Su |
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