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 added value of internal audit greatly contributes to adding value to the institution, but most departments of economic units in Iraq neglected the role of internal audit and the added value that can be achieved by those institutions, since the term added value of internal audit is a relatively vague term from the premise that what cannot be measured is difficult Determine it, and perhaps descriptive standards for it is the extent of compliance with international auditing standards (IIA).
The research aims to study the procedures and results of auditing to verify that they have given an added value to the audit with a positive impact, develop its aspects and research, identify deficiencies for the audi
... Show MoreThe [2-hydroxy-1, 2-diphynel-ethanone oxime] was reacted with 1, 2-dichloroethan to give the new ligand [H2L]. this ligand was reacted with some metal ions (Co (II), Ni (II), Cu (II), Zn (II) and Cd (II) in methanol as a solvent to give a series of new (1: 1) complexes of the general formula [M (HL)] Cl,(where: M= Co (II), Ni (II), Cu (II), Zn (II) and Cd (II)) are isolated All compounds have been characterized by spectroscopic methods [IR, UV-Vis] atomic absorption. Chloride content along with conductivity measurements. From the above data the proposed molecular structure for (Co, Cu, Ni, Zn and Cd) complexes adopting a tetrahedral structure
The Vulnerable Indian Roofed Turtle Pangshura tecta (Gray, 1831) (Testudines: Geoemydidae) occurs in the Sub-Himalayan lowlands of India, Nepal, Bangladesh, and Pakistan. Little is known about its natural history, no studies have been conducted revealing its natural predators. In this study, a group of Large-billed Crow Corvus macrorhynchos Wagler, 1827 (Passeriformes: Corvidae) was observed hunting and predating on an Indian Roofed Turtle carcass in the bank of river Kuakhai, Bhubaneswar, India. The first record of this predation behaviour is reported and substantiated by photographic evidence.
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Abstract
This Research aims for harnessing critical and innovative thinking approaches besides innovative problem solving tools in pursuing continual quality improvement initiatives for the benefit of achieving operations results effectively in water treatment plants in Baghdad Water Authority. Case study has been used in fulfilling this research in the sadr city water treatment plant, which was chosen as a study sample as it facilitates describing and analyzing its current operational situation, collecting and analyzing its own data, in order to get its own desired improvement opportunity be done. Many statistical means and visual thinking promoting methods has been used to fulfill research task.
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Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is a potent ligand for AhR and a known carcinogen. While AhR activation by TCDD leads to significant immunosuppression, how this translates into carcinogenic signal is unclear. Recently, we demonstrated that activation of AhR by TCDD in naïve C57BL6 mice leads to massive induction of myeloid derived-suppressor cells (MDSCs). In the current study, we investigated the role of the gut microbiota in TCDD-mediated MDSC induction. TCDD caused significant alterations in the gut microbiome, such as increases in Prevotella and Lactobacillus, while decreasing Sutterella and Bacteroides. Fecal transplants from TCDD-treated
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The research aims to identify the effect of the training program that is based on integrating futuristic thinking skills with classroom interaction patterns on mathematics teachers in order to provide their students with creative solution skills. The research sample consisted of 31teachers (15 teachers for the experimental group and 16 for the control groups). The researcher developed a measure for the academic self-efficacy consisting of (39) items. Its validity, reliability, coefficient of difficulty and discriminatory power were estimated. To analyze the findings, the researcher adopted the Mann-Whitney (U) test and the effect size, and the findings were as follows: There is a statistically significant difference at the significance leve
... Show MoreTwo Schiff bases, namely, 3-(benzylidene amino) -2-thioxo-6-methyl 2,5-dihydropyrimidine-4(3H)-one (LS])and 3-(benzylidene amino)-6-methyl pyrimidine 4(3H, 5H)-dione(LA)as chelating ligands), were used to prepare some complexes of Cr(III), La(III), and Ce(III)] ions. Standard physico-chemical procedures including metal analysis M%, element microanalysis (C.H.N.S) , magnetic susceptibility, conductometric measurements, FT-IR and UV-visible Spectra were used to identify Metal (III) complexes and Schiff bases (LS) and (LA). According to findings, a [Cr(III) complex] showed six coordinated octahedral geometry, while [La(III), and Ce(III) complexes]were structured with coordination number seven. Schiff's bases a
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Erratum for Organic acid concentration thresholds for ageing of carbonate minerals: Implications for CO2 trapping/storage.
The research has deal with the relationship between organizational justice and empowerment and their impact on the achievement of organizational commitment in the office of Labour and Vocational Training. To study the research problem which is represented a sense that employees with low levels of organizational justice and empowerment and the reflection on the organizational commitment of the employees, so that Has been collecting data and information relating to research by designing a questionnaire, were distributed to a sample of (50) people in the office mentioned, and the results of the study to confirm the research hypotheses. and the key results of the research was the presence of correlation relationships and the effect o
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