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.
Copper with different concentrations doped with zinc oxide nanoparticles were prepared from a mixture of zinc acetate and copper acetate with sodium hydroxide in aqueous solution. The structure of the prepared samples was done by X-ray diffraction, atomic force microscopy (AFM) and UV-VIS absorption spectrophotometer. Debye-Scherer formula was used to calculate the size of the prepared samples. The band gap of the nanoparticle ZnO was determined by using UV-VIS optical spectroscopy.
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Abstract
The current research aims to clarify the role of local policies on the performance of the province of Baghdad, after studies proved practical experience what those policies of the major role and effect on the lives of citizens, as well as alleviate the burden on central government, which make a lot of states give local governments broad powers and her specialty funds for the exercise of its vital role and actor in various joints of local development, research has indentified a problem in a number of questions such as: do you have the policy of the provincial council of local qualified and able to influence the performance of the province? What are the main forces of powerful and implementation of policies at all
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According to the prevalence of multidrug resistance bacteria, especially Pseudomonas aeruginosa, in which the essential mechanism of drug resistance is the ability to possess an efflux pump by which extrusion of antimicrobial agents usually occurs, this study aims to detect the presence of mexB multidrug efflux gene in some local isolates of this bacteria that show resistance towards three antibiotics, out of five. Sensitivity test to antibiotics was performed on all isolates by using meropenem (10μg/disc), imipenem (10μg/disc), amikacin (30 μg/disc), ciprofloxacin (5μg/disc) and ceftazidime (30 μg/disc). Conventional PCR results showed the presence of mexB gene (244bp) in four isolates out of ten (40%). In addition,25, 50μg/ml of cur
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Dermatophytes are species with slight genetic variation, and are yet several uncertainties about the differences among species. This study aims to isolate and diagnose the Trichophyton interdigitale by molecular technique and to reveal the phylogenetic distance and similarity of the Iraqi isolates to other isolates from the globe, in addition, to submit the obtained sequences to the NCBI database. This study included 86 with multiple lesions on different parts of the body. The results showed different variations within the ITS gene between the isolates. It was concluded that Trichophyton interdigitale in Iraqi isolates had two types of substitution variations (Transition and Transversion) different than global isolates. Moreover, it
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Traffic loading and environmental factors are among the most serious variables that cause the spoilage of flexible pavements and lead to a decrease in their design life. The objective of this study is to investigate the influence of axle load raise and the change in resilient modulus on the flexible pavement design life. Locally, Highway geometric design code for Iraqi building code has assign certain admissible maximum load limits per every axle truck type that should not be overrun. In this paper nine different axle truck loads (8, 9, 10, 11, 12, 13, 14, 15, and 16) tons, single axle with dual tire and, and two different resilient moduli of asphalt pavement were chosen. The evaluation was carried out assuming high temperature to represent
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In this research study theory to find the stress and emotion gases in the glass as a result of exposure to pulses of the laser beam has been the study using vehicles three major on-system axes cylindrical (r, 0, z), where I took three models of glass silica glass soda glass fused and shedtwo types of lasers where the study showed that the thermal stresses and emotions ...
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ZnIn2(Se1-xTex)4 (ZIST) chalcopyrite semiconductor thin films at various contents (x = 0.0, 0.2, and 0.4) are deposited on glass and p type silicon (111) substrate to produce heterojunction solar cell by using the thermal evaporation technique at RT where the thickness of 500 nm with a vacuum of 1×10-5 mbar and a deposited rates of 5.1 nm/s. This study focuses on how differing x content effect on the factors affecting the solar cell characteristics of ZIST thin film and n-ZIST/p-Si heterojunction. X-ray diffraction XRD investigation shows that this structure of ZIST film is polycrystalline and tetragonal, with (112) preferred orientation at 2θ ≈ 27.01. Moreover, atomic force microscopy AFM is studying the external morphology of
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