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 present study investigated the impact of fuel kind on the emitted emissions at the idling period. Three types of available fuels in Iraq were tested. The tests conducted on ordinary gasoline with an octane number of 82, premium gasoline with an octane number of 92, and M20 (consist of 20% methanol and 80% regular gasoline). The 2 liters Mercedes-Benz engine was used in the experiments.
The results showed that engine operation at idle speed emits high levels of CO, CO2, HC, NOx and noise. The produced emission levels depend highly on fuel type. The premium gasoline (ON=92) represents the lower emissions level except for noise at all idling speed. Adding methanol to ordinary gasoline (ON=82) showed high levels of emi
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Praise be to God, Lord of the worlds, and peace and blessings be upon our master Muhammad, and upon his whole family and companions, but after: God Almighty has instituted types of acts of worship and classes of obedience that, if performed by the servant, would link him with his Creator and connect with his Lord. And for the sake of these worship, and the greatest of them: the worship of i'tikaaf, for souls need rest and recuperation from the struggle of life and its concerns, concerns and anguish, and people differ in their attitudes and interests in the means of recreation, and many people consider the outing and travel the best way in which they breathe from their toil and toil, and reclaim their strength and the advised adult He fin
... Show MoreThe potential application of granules of brick waste (GBW) as a low-cost sorbent for removal of Ni+2ions from aqueous solutions has been studied. The properties of GBW were determined through several tests such as X-Ray diffraction (XRD), Energy dispersive X-ray (EDX), Scanning electron microscopy (SEM), and BET surface area. In batch tests, the influence of several operating parameters including contact time, initial concentration, agitation speed, and the dose of GBW was investigated. The best values of these parameters that provided maximum removal efficiency of nickel (39.4%) were 1.5 hr, 50 mg/L, 250 rpm, and 1.8 g/100mL, respectively. The adsorption data obtained by batch experiments subjected to the Three i
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This study is to investigate the possibility of using activated carbon prepared from Iraqi date-pits (ADP) which are produced from palm trees (Phoenix dactylifera L.) as low-cost reactive material in the permeable reactive barrier (PRB) for treating lead (Pb<sup>+2</sup>) from the contaminated groundwater, and then compare the results experimentally with other common reactive materials such as commercial activated carbon (CAC), zeolite pellets (ZP). Factors influencing sorption such as contact time, initial pH of the solution, sorbent dosage, agitation speed, and initial lead concentration has been studied. Two isotherm models were used for the description of sorption data (Langmuir and Freundlich). The maximum lead sorp
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This research aims at identifying the commitment of satellite news channels in Arabic to the set of important standards that reflect their credibility in dealing with the media material, and considering that these channels give special importance to events in Iraq, as well as the Arab region and the world, decide to choose them and study them with a problem The research was a question about the level of credibility of Iraqi media. This research is descriptive research, which used the survey method on an objective sample of 245 items, while the questionnaire was used as a data collection tool. Seven channels were selected in Arabic for the study. The three most watched channels were chosen. These channels included the channels of Russia t
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A survey to show the reality of educational guidance in schools in Baghdad Rusafa 3\ from the point of view of educational guides. It used the analytical descriptive technique. The research community consists of () male and female advisors in schools of Baghdad. The sample was haphazard( 51\.) The questionnaire was made and it consists (50) items. The research statistics have been analyzed by (SPSS), The questionnaires have been used in the first term in (2017-2018) and the results showed the relationship between the advisor and other teachers, the advisor and students fathers and the local society. According to the results, the research made some recommendations and suggestions.
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A new optoelectronic flow injection method is proposed for the determination of ferrous ions (Fe 2+ ) based on thiocyanate complexation to form a deep-red FeSCN 2+ complex.
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