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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The -mixing of - transition in Er 168 populated in Er)n,n(Er 168168 reaction is calculated in the present work by using a2- ratio method. This method has used in previou studies [4, 5, 6, 7] in case that the second transition is pure or for that transition which can be considered as pure only, but in one work we applied this method for two cases, in the first one for pure transition and in the 2nd one for non pure transitions. We take into accunt the experimental a2- coefficient for p revious works and -values for one transition only [1]. The results obtained are, in general, in agood agreement within associated errors, with those reported previously [1], the discrepancies that occur are due to inaccuracies existing
... Show Moreتضمن هذا العمل تحضير ليكند قاعدة شيف جديدة مشتقة من مادة البولي أكريلاميد والكلوترالديهايد [(2S, 2'S) – N, N' - (pentane-1, 5-diylidene) bis (2- methylbutan amide)] مع بعض المعادن الثقيلة (Cr + 3 Mn + 3 , Fe + 3 , ,Co + 2, Ni + 2 ,Cu + 2 Zn + 2 , Cd + 2,) لتنتج المعقدات المقابلة. تم تشخيص قواعد شيف ومعقداتها المعدنية بأستخدام طيف الأشعة تحت الحمراء والأشعة المرئية وفوق البنفسجية، والتوصيلية ,وقيم المغناطيسية والتحليل الحراري الوزني وحيود الأشعة السينية ومجه
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Freshwater mussels are a guild of stationary, suspended-feeding species; they perform significant ecological functions like nitrogen cycling, bioturbation that gives oxygen and habitat that other creatures need to survive, and increasing water clearance by filtration. Knowledge of the freshwater mussel Unio tigridis Bourguignat, 1852, distribution, and molecular study in Iraq was inadequate. In the current study, this species of freshwater Mussels belonging to the family Unionidae was collected from different locations in the Greater Zab River, from April 2022 to November 2022. The average water temperature of the site was arranged between (17.8 to 36.1 C°). All previous studies in the Kurdistan Region and Iraq were based on morphologic
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This research represents a practical attempt applied to calibrate and verify a hydraulic model for the Blue Nile River. The calibration procedures are performed using the observed data for a previous period and comparing them with the calibration results while verification requirements are achieved with the application of the observed data for another future period and comparing them with the verification results. The study objective covered a relationship of the river terrain with the distance between the assumed points of the dam failures along the river length. The computed model values and the observed data should conform to the theoretical analysis and the overall verification performance of the model by comparing i
... Show MoreThe remove of direct blue (DB71) anionic dye on flint clay in aqueous solution was investigated by using a batch system for various dye concentrations. The contact time, pH, adsorbent dose, and temperature was studied under batch adsorption technique. The data of adsorption equilibrium fit with isotherm Langmuar and Freiundlich ,when the correlation coefficient used to elucidate the best fitting isotherm model. The thermodynamic parameters such as, ?Hº ,?Sº and ?Gº. Thermodynamic analysis indicated that the sorption of the dyes onto Flint clay was endothermic and spontaneous.
Background: Esthetic treatment is the options of patient seeking orthodontic treatment. Therefore this study was conducted to measure the concentration of Aluminum, Nickel, Chromium and Iron ions released from combination of monocrysralline brackets with different arch wires immersed in artificial saliva at different duration, to evaluate the corrosion point on different parts of the orthodontic appliances before and after immersion in artificial saliva, and to evaluate the corrosion potential of each group of the orthodontic appliances. Material and methods: Eighty orthodontic sets prepared. Each set represents half fixed orthodontic appliance, from the central incisor to the first molar, for the maxillary arch, each set consisted of molar
... Show MoreThe survey was carried out From January to April of 2018 on macrofungi samples collected from different places in Halabja province located in north eastern parts of Iraq-Kurdistan region. This region is rich in forest trees and pasture lands with diversity of shrubs and herbs and is expected to support the growth of several macro fungal species. However, this part of Kurdistan in Iraq is still unexplored from macrofungal point of view. In this paper three species from Pezizaceae and Pyronemataceae families that belonging to (Pezizales, Ascomycota), were reported from Iraqi Kurdistan. These macrofungal species are recorded for the first time from Iraq. Also the species were identified and showing their locations distributed on a map prepared
... Show MoreThe present work aimed to study the efficiency of nanofiltration (NF) and reverse osmosis (RO) process for water recovery from electroplating wastewater and study the factors affecting the performance of two membrane processes. Nanofiltration and reverse osmosis membranes are made from polyamide as spiral wound module. The inorganic materials ZnCl2, CuCl2.2H2O, NiCl2.6H2O and CrCl3.6H2O were used as feed solutions. The operating parameters studied were: operating time, feed concentrations for heavy metal ions, operating pressure, feed flow rate, feed temperature and feed pH. The experimental results showed, the permeate concentration increased and water flux decreased with increase in time from 0 to 70 min. The permeate concentrations incre
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