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.
Arts acts to reduce and exclude everything that is not necessary in the delivery of meaning, especially representative performance, which is based on the intensification of the physical and vocal actions, and in drawing the dimensions of the character. And because that, the artistic staff, among others in the ministry of education, are considered the cornerstone, in the development of theatrical activities in schools, this research came to find out the extent to which teachers in primary schools, in secondary schools and artistic supervisors rely on shorthand mechanisms in the representative performance.
The current research relied on the descriptive approach, in analyzing his sample, which was deliberately chosen, Among the most imp
Vorwort:
Die Übersetzung gilt in der Vergangenheit als Brücke zwischen verschiedenen Gesellschaften. Sie spielt eine große Rolle im Bereich Kulturaustausch der Länder. Die Araber haben das früh betrachtet, und die Übersetzung als Wissenschaft wird eine der wichtigsten Lehren. Viele griechische Wissenschaften waren durch Übersetzung in den arabischen Ländern bekannt.
In dieser Forschung findet man einen Überblick über die Wichtigkeit der Sprichwörterübersetzung und welche Rolle spielt diese Übersetzung im Z
... Show MoreThis research sheds light on the morphological structure of nouns and verbs in the novel "ASTONISHMENT" by the novelist Aharon Applefield by analysing selected models from the novel in a morphological analysis in order to identify the most important morphological features of this structure according to a statistical analytical approach.
The morphological structure is the main pillar of the linguistic structure of the literary text. Morphology is the science that studies the word, by which its structure and original letters are known, and the change that has occurred to it. The aesthetics of the fictional text is reflected in this structure, through which the writer conveys his ideas and narrations to the reader. The research
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This work focuses on the preparation of pure nanocrystalline SnO2 and SnO2:Cu thin films on cleaned glass substrates utilizing a sol-gel spin coating and chemical bath deposition (CBD) procedures. The primary aim of this study is to investigate the possible use of these thin films in the context of gas sensor applications. The films underwent annealing in an air environment at a temperature of 500 ◦C for duration of 60 minutes. The thickness of the film that was deposited may be estimated to be around 300 nm. The investigation included an examination of the structural, optical, electrical, and sensing characteristics, which were explored across various preparation circumstances, specifically focusing on varied
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Objective : Sciatic nerve block (popliteal approach) and femoral N block is a new technique other than general anesthesia in below knee surgery because it provides adequate muscle relaxation, with good intraoperative and post-operative analgesia. Nefopam is non opioid, non-respiratory depressant and non-sedative was mixed with local anesthetics drug to study the effects. This study was done to compare the onset and duration of sensory and onset time and duration of action of motor block following administration of either bupivacaine alone with administration of bupivacaine and Nefopam in patients undergoing below knee lower limb surgeries under ultrasound guided regional anesthesia.
Methods: 100 patients with American society of anest
Addressed this research the impact of intelligence emotional dimensions of the main(self awareness, and self-management, and social awareness, and relationship management) in the performance excellence the university(performance optimization, and strategic development) this is by middling the styles decision making which are (rational and intuitive, and dependent, and spontaneous, and avoidant), and Go search of an intellectual dilemma raise fundamental questions revolve around the search was to answer those questions through a theoretical framework for search variables first and test models of the relationship and second through the impact six hypotheses President.The objective of the research to make sure the contr
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The research includes the synthesis and identification of the mixed ligands complexes of M(II) Ions in general composition [M(Lyn)2(phen)] Where L- lysine (C6H14N2O2) commonly abbreviated (LynH) as a primary ligand and 1,10-phenanthroline(C12H8N2) commonly abbreviated as "phen," as a secondary ligand . The ligands and the metal chlorides were brought in to reaction at room temperature in ethanol as solvent. The reaction required the following molar ratio [(1:1:2) (metal): phen:2 Lyn -] with M(II) ions, were M = Mn(II),Cu(II), Ni(II), Co(II), Fe(II) and Cd(II). Our research also includes studying the bio–activity of the some complexes prepared against pathogenic bacteria Escherichia coli(-),Staphylococcus(-) , Pseudomonas (-), Bacillus (-)
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