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.
This study was conducted to determine the ability of water treatment system (Vortisand) to reduce some chemical and physical properties for tigris river raw water, It consisted of turbidity, electrical conductivity, pH, total hardness, calcium Hardness as well as temperature in order to determine the unit`s efficiency for reducing their concentration as compared to those in the water produced by some classical potable water projects (Dora and Wathba) in Baghdad. Samples were collected during the cold months (December 2016 and January 2017) and during the hot months (May and June 2017). The results showed that this system has the ability to reduce some properties such as turbidity, the values were 215NTU in raw water and decreased to NTU
... Show MoreBackground: Obesity typically results from a variety of causes and factors which contribute, genetics included, and style of living choices, and described as excessive body fat accumulation of body fat lead to excessive body, is a chronic disorder that combines pathogenic environmental and genetic factors. So, the current study objective was to investigate the of the FTO gene rs9939609 polymorphism and the obesity risk. Explaining the relationship between fat mass and obesity-associated gene (FTO) rs9939609 polymorphism and obesity in adults. Methods: Identify research exploring the association between the obesity risk and the variation polymorphisms of FTO gene rs9939609. We combined the modified odds ratios (OR) as total groups and subgro
... Show MoreRKRAS L. K. Abdul Karem, F. H. Ganim, Biochemical and Cellular Archives, 2018 - Cited by 2
MJ Abbas, AK Hussein, Journal of Physical Education, 2019
The special core analysis tests were accomplished on a set of core plugs for Mishrif Formation (mA, mB1, and mB2cde/mC units) in West Qurna/1 oilfield, southern Iraq. Oil relative permeability (Kro) data and the Corey-type fit of the data as functions of the brine saturation at the core outlet face for individual samples in the water-oil imbibition process to estimate relative permeability measurements by the centrifuge method were utilized. Identical correlations for oil and water relative permeabilities were extracted by steady-state and unsteady-state methods. For the mA samples, the gas-water capillary pressure curves were within a narrow range (almost identical) indicating that mA is a homogeneous unit. Kro curves for three mB2
... Show MoreA simple, sensitive and rapid method was used for the estimate of: Propranolol with Bi (III) to prove the efficiency, reliability and repeatability of the long distance chasing photometer (NAG-ADF-300-2) using continuous flow injection analysis. The method is based on a reaction between propranolol and Bi (III) in an aqueous medium to obtain a yellow precipitate. Optimum parameters were studied to increase the sensitivity for the developed method. A linear range for calibration graph was 0.1-25 mmol/L for cell A and 1-40 mmol/L for cell B, and LOD 51.8698 ng/200 µL and 363.0886 ng /200 µL , respectively to cell A and cell B with correlation coefficient (r) 0.9975 for cell A, 0.9966 for cell B, RSD% was lower than 1%, (n = 8) for the
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