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 tunnel’s stability during construction is a very important matter. Some methods have been proposed for stability evaluation, but the hazard warning levels (HWLs) are more applicable among these methods. Despite monitoring and applying HWLs, several collapses in Shibli twin tunnels in Iran have cast doubts on the accuracy of this criterion in the presence of water. In this study, the critical strains under different water contents were measured through uniaxial compressive strength tests on 11 different shale and marl samples. A comparison of laboratory tests and numerical results shows that the influence of the moisture content on the critical strain is negligible. In addition, the results show that there is no dir
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The issue of sports media and its role in winning sports and non-sports fans and unifying them towards various issues and events is one of the important topics that have begun to occupy many countries and governments in the world and allocate appropriate spaces for it and provide it with sufficient funds and budgets to advance this reality...
Especially since the concept of the sports official has It changed a lot and turned into an economic concept rather than a sport one through the investment in the field of sports and the entry of capital and contributes to the prosperity of the countries interested in this field.
It is important that includes profit and loss in addition to knowing how to employ and win the masses and e
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Production and characterization of methionine γ- lyase from Pseudomonas putida and its effect on cancer cell lines
In this paper, we consider a new approach to solve type of partial differential equation by using coupled Laplace transformation with decomposition method to find the exact solution for non–linear non–homogenous equation with initial conditions. The reliability for suggested approach illustrated by solving model equations such as second order linear and nonlinear Klein–Gordon equation. The application results show the efficiency and ability for suggested approach.
The study aimed to evaluating the inhibitory activity of apigenin extracted from Salvia officinalis leaves on the growth of L20B cancer cell in vitro, and through two incubation periods; 48 and 72 hours. Accordingly, eight concentrations (1.56, 3.13, 6.25, 12.5, 25.0, 50.0, 100.0 and 200.0 micromol) of apigenin and similar concentrations of vitamin C and carbon tetrachloride (CCl4) were tested. The apigenin revealed its significant inhibitory potentials against the growth of L20B cell line, especially at the low concentrations (1.56, 3.13 and 6.25 micromol) and at 72 incubation period in comparison with vitamin C and CCl4.
Giant Cell Fibroma (GCF) is a relatively rare oral mucosal lesion, so named due to the characteristic giant cells present within the fibrous stroma of the lesion, limited number of clinicopathological studies were performed in previously published literature. This study was performed to evaluate the clinicopathological features of Giant cell fibroma in a sample of Iraqi patients. Formalin-fixed paraffin-embedded sections from 22 giant cell fibroma in period between 2010 and 2018 were retrieved from the laboratory of oral pathology of Baghdad University/College of Dentistry, Clinical data and microscopic features were reviewed and analyzed according to the available surgical reports. The mean age of patients at the time of diagnosis was 29.6
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Visceral leishmaniasis (VL) or kala-azar is one of the worlds most neglected tropical diseases in mortality and fourth in morbidity, rK39 dipstick was used to diagnose the suspected infected patients as cheapest simple technique which can differentiate recent from chronic infection, for disease out-coming, naïve T-lymphocyte cells should be differentiated into pathogen-specific immunity responses, such as T-helper 1(Th-1) or (Th-2). HLA-G is a special protein defined as nonclassical HLA class I molecule can suppress the immune system through prevention of T-cell function by foul all T-cell mechanisms. So, this study aimed to detect and evaluate the level of sHLA-G in the sera of patients infected with VL. The results showed that there was
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