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.
The purpose of the current research is to identify the most important problems that primary school students suffer from inside and outside the classroom from the point of view of their teachers. A sample of (100) male and female teachers was chosen from the Rusafa\ second Directorate for the academic year (2018-2019). The research tool was prepared after reviewing literature related to the issue of problems and difficulties facing students or students in the school stage and even at university. The researcher reached several results that were discussed in the fourth chapter, with a set of conclusions based on the results of the research, and come up with several recommendations and suggestions.
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The letters of the Al-Safa Brothers contain many tales, in which each story is a literary text that is independent from the other texts, but it constitutes a unified entity in its entirety, because these stories contain recurring structural elements that go into building each one of them. The stories - the subject of the analysis - were not keen on something that was keen on adhering to the Sufi vision and its apparent effect on the tales of the messages, starting with the announcement of the narration to the narrator and his attempt to persuade the recipient of the truth of what is being told and its realism, and made him believe it. The stories correspond to the formal construction of the ideal style defined by th
... Show MoreThis study deals with the conceptual rooting of the practical relationship between the practice of international public relations and popular diplomacy, as the latter sought - as a legitimate extension of traditional diplomacy - to involve foreign residents in international public relations capable of drawing a positive image of the state among people of other countries, by employing modern communication technology. Especially social media, which led to a shift from old diplomacy to digital public diplomacy. The research paper seeks to analyze the communicative content of the Facebook page (Israel in the Iraqi dialect), which a page is belonging to the "Israeli" Ministry of Foreign Affairs that aims, according to its messages, to encoura
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Some of the characters of the Staphylolysin A and D enzymes purified from Pseudomonas aeruginosa P16 and P5 respectively were studied, the molecular weights of Staphylolysin A and D were 20.417 kilo dalton and 23.988 kilo Dalton respectively by SDS- polyacryl amide gel electrophoresis. The optimum pH for staphylolysin A activity was found to be 8 which gives higher activity reaches 150 unit/ml, and for enzyme stability was 7.5-8.5 in which the enzyme nearly retained its full activity, while it was 9.5 for staphylolysin D that gives higher activity of 16 unit/ml,and 8.5-9.5 for enzyme stability in which the enzyme nearly retained its full activity, Maximum activity of two enzymes was obtained at 40C in which the specific activity for st
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The present work represents description of three new species of genus Anthrenus
Geoffory from Iraq, these are : A. aradensis sp. nov., A. fabrici sp. nov. and A.
unicolor sp. nov. Locality, host plants and date of collection were given.
Pyrolysis of virgin polyethylene plastics was studied in order to produce hydrocarbon liquid fuel. The pyrolysis process carried out for low and high-density polyethylene plastics in open system batch reactor in temperature range of 370 to 450°C.
Thermo-gravimetric analysis of the virgin plastics showed that the degradation ranges were between 326 and 495 °C. The results showed that the optimum temperature range of pyrolysis of polyethylene plastics that gives highest liquid yield (with specific gravity between 0.7844 and 0.7865) was 390 to 410 °C with reaction time of about 35 minutes. Fourier Transform Infrared spectroscopy gave a quite evidence that the produced hydrocarbon liquid fuel consisted ma
... Show Morekindergarten teacher is one of the fundamental pillars upon which the kindergarten environment so that exposure to a number of problems that could affect its functioning in addition to any deficiencies in this environment leads to deprive a child of some activities and acquisition of concepts so the researcher studying the problems of working in kindergartens from the perspective of the parameters, so the researcher based measuring instrument for labour problems of (30) search sample was paragraph (50) parameter that was chosen at random and have been extracted Sincerity and strength tool researcher used statistical methods and discriminatory (Pearson correlation coefficient, t
... Show MoreThis research has tackled a scientific figure belongs to Koofi grammatical school. He is Abdullah Al-Tuaal who has occupied the fourth layer among the Koofi grammarians. There was a confusion between him and another Koofi grammarian from the same layer (Mohammed Bin Qaadim) in some old references. This research has clarified this confusion to affirm that they are two different grammarians.This research also tackles the scientific status of Al-Tuaal and the grammatical views that have been ascribed to him in addition to the analysis and discussion of these views. All this in the light of what has been mentioned in grammar books about the issues in which these views were stated.