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.
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research summary
Praise be to Allah، and prayers and peace be upon His Messenger and His chosen one، his family، companions and his family and his family.
After that، this is a study on the question of the claim، as one of the controversial questions that are the mainstay of debates، and I looked at it: the limit of the question of the claim، its importance، its impact، its aspects of corruption، its sections، and the representation of its sections of the fundamentalist controversy;
The research was according to the following plan:
Introduction، in which she mentioned: the importance of the research topic and the reason for its selection، its questions، its l
... Show MoreThis research was conducted in order to monitor and measure the dimensions of media policy in satellite channels directed from the point of view of the communicator, and this research is classified among the descriptive studies, as the researcher used the survey method to answer the questions that were formulated in light of the research problem represented by the main question: What are the dimensions of media policy in Directed satellite channels? .
To achieve the objectives of the study, the researcher used the following tools:
The questionnaire, in order to survey the attitudes of communicators about the extent to which the media policy during crises reflects on their professional standards. The research community is represente
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The research aims to assess the claystone exposed in the Nfayil Formation (Middle Miocene) for Portland cement (P.C.) manufacturing based on mineralogy and geochemistry. The importance of the study is to avoid the miming of the agricultural soils that are mining now for the cement industry. Claystones of Nfayil Formation and the limestone of the Euphrates Formation were used to design the raw mixture as clay to limestone (1:3). The chemical composition (%) of the designed mixture was calculated using the Alligation Alternative Method (A.A.M.) as CaO (65.52), MgO (1.05), SiO2 (21.65), Al2O3 (7.43), Fe2O3 (2.62), Na2O3+K2O (1.52) and SO3 (0.26), which are suitable for P.C. The lime saturation factor (LSF = 92.8), silica saturation fac
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Arsenic is a prevalent and pervasive environmental contaminant with varied amounts in drinking water. Arsenic exposure causes cancer, cardiovascular, liver, nerve, and ophthalmic diseases. The current study aimed to find the best conditions for eliminating arsenic from simulated wastewater and their effect on biomarkers of hepatic in mice. Adsorption tests including pH, contact duration, Al-kheriat dosage, and arsenic concentrations were evaluated. Seventy-two healthy albino mice (male) were accidentally allocated into nine groups (n = 8), the first group was considered as healthy control, the second group (AL-Kheriat), and other groups received AL-Kheriat and arsenic 25, 50, 75, 100, 125, 150 and 175 mg/kg, respectively. Next 10 days, the
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The aim of the research is to:. Preparation and implementation of special educational units using multimedia to learn the skill of scrolling from below. 2 to recognize the impact of the use of multimedia in learning the skill of scrolling from below. 3 to identify the differences between the tests after the two groups research in learning the skill of passing from the bottom volleyball. The research represented the students of the first stage and the sample of the research was drawn randomly and the number of (50) students were divided into two experimental and control groups and each group (25) students were used standardized tests and conducting pre-tests and the implementation of the main exp
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The study aims to demonstrate the significance of metaverse technology across various disciplines, academic degrees, scientific fields, and academic titles. It also aims to assess the level of knowledge and understanding of university teachers (research sample) regarding metaverse technology. Hence, the descriptive research methodology was based on the method of statistical survey in the sample. It involved a set of organized scientific steps to deduce data from the reality of the statistical sample and its nature in order to achieve the objectives of the study. In this study, a questionnaire was used as a tool to collect data from a random sample of approximately 121 teachers and instructors from the University of Baghdad. This app
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This investigation showed (31) species belonging to (15) genera under (five) families and two orders. The leafminers Dipter families (Agromozidae, Anthomyiidae, Drosophilidae), Agromyzid flies is the highest level of investigated many host plants, but other families have lowest host plants. The synonyms of species were provided from GBIF scarlet's. The date and localities of sampling collection were recorded.
In this research, Schiff bases derived from the reaction of anthrone with different heterocyclic amines have been described. The resulted Schiff base compounds were reacted with various nucleophiles in order to obtain new heterocyclic derivatives. Chemical structures of all products were confirmed by IR, 1H-, 13C-NMR spectral data and elemental analysis. All synthesized compounds were in vitro tested against a standard strain of pathogenic microorganism including Gram +ve bacteria (Staphylococcus aureus), Gram –ve bacteria (Escherichia coli), and fungi (Candida albicans).
Soils at Al-Koot-Btera were choosen to determine their sedimentary
environments. It is found that there are , five soil series and as mentioned :
MF11-MW9-DM97-DM57-DF95 . The five found soil series are of internal
well drained ,moderate and imperfect. Their textures vary in moderately,fine
and moderately fine.
indicating that sediments rang from poorly to very poorly sorting. Values of median
diameter Md Ø range between 4.11-7.80 Ø .The relation between the sorting
and median diameter shows that 95.24% of samples is a sedimentary
environment of aquite river , while 4.76% is aeolian sediments.
The values of meso to platy kurtic of most horizon materials rang
between 0.67-1.26 Ø .That is to say the samples