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 research aims to identify the relationship between employing future skills during teaching from the viewpoint of students of Islamic studies at the Northern Border University, as well as their attitudes towards future professions. The researcher employed the correlational descriptive approach. The tools were a questionnaire for employing future skills, and a scale for the attitude towards the future profession. The two research tools were applied to a random sample of (242) male and female students from the department of Islamic Studies, College of Education and Arts. The findings showed that the total level of employing future skills and their three axes during teaching was average. It was also found that the attitude towards future
... Show MoreThis study aims to identify the degree of students of Princess Rahma University College owning e-learning skills related to MOODLE as they perceived in the of light Corona crisis. The researchers' questionnaire consisted of (37) items, distributed in three areas of e-learning skills related to the MOODLE on (147) students were chosen randomly. The results of the study showed that the degree of students 'possession of e-learning skills related to the MOODLE was significant. The results also revealed that there were statistically significant differences in the degree of students' possession of electronic learning skills related to the MOODLE due to sex in favor of females. Finally, there were no statistically significant differences in the
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Nowadays, university education stands in front of both students who feel they are weak and teachers who are addicted to using traditional and dependent teaching. This has led to have negative repercussions on the learner from different aspects, including the mental aspect and the academic achievement process. Therefore, the present research is concerned with finding a new teaching method that adopts the motivation by the fear of failure technique. Thus, the study aims to examine the effect of adopting this method on students’ academic achievement. To achieve this aim, an experimental method was used, and an achievement test was built for the curriculum material of level two students. The pretest test was applied on 17 male and female s
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Water scarcity is one of the most important problems facing humanity in various fields such as economics, industry, agriculture, and tourism. This may push people to use low-quality water like industrial-wastewater. The application of some chemical compounds to get rid of heavy metals such as cadmium is an environmentally harmful approach. It is well-known that heavy metals as cadmium may induce harmful problems when present in water and invade to soil, plants and food chain of a human being. In this case, man will be forced to use the low quality water in irrigation. Application of natural materials instead of chemicals to remove cadmium from polluted water is an environmental friendly approach. Attention was drawn in this research wor
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A mounted specimen of a mustelid animal deposited in the Kurdistan Museum of Natural History, Salahaddin University, Erbil proved to be Mustela erminea (Linnaeus, 1758) and represents a new record for the mammalian fauna of Iraq. Its measurements and some biological noted are provided. Also, two passerine birds; the Red-headed bunting, Emberiza bruniceps Brandt, 1841(Family, Emberizidae) and the Variable wheatear, Oenanthe picata (Blyth, 1847) (Family, Muscicapidae) were recorded for the first time in Iraq. Furthermore, the tree frog Hyla savignyi Audouin, 1829 was found in two locations north east of Iraq with spotted dorsum and having interesting behavior in having the capabil
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This work studies the performance of zeolite permeable reactive barrier in removing cadmium from a contaminated shallow aquifer. Batch tests have been performed to characterize the equilibrium sorption properties of the zeolite in cadmium-containing aqueous solutions. A 1D numerical finite difference model has been developed to describe pollutant transport within groundwater taking pollutant sorption on the permeable reactive barrier (PRB), which is performed by Langmuir equation, into account. Numerical results show that the PRB starts to saturate after a period of time (~120 h) due to reduction of the retardation factor, indicating a decrease in the percentage of zeolite functionality. However, a reasonable agreement between model predict
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