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 current study aims at using non-hatchable artemia eggs of local origin and making use of these eggs by decapsulating and presenting them as food for the larvae of the Cyprinus carpio as a source of animal protein with high nutritional value instead of throwing them away. The results showed that the second parameter (A2) was highly significant at the level (P≤0.05) in the growth rates of the larvae that were fed on decapsulated artemia eggs alone, and it was better than the two control parameters (A1), in which the larvae were fed with feed designated for Cyprinus carpio fish. It also outperformed the third parameter (A3), in which the feed was mixed with artemia eggs with 50% decapsulation, which also outperformed the control paramete
... Show MoreThe current research aims to shed light on the Global Reporting Initiative (GRI), which helps to report financial and non-financial information by economic units in general and listed on the Iraq Stock Exchange in particular. The research was based on a main premise that apply the criteria of the Global Reporting Initiative (GRI) would provide useful information to users to help them make appropriate decisions. To achieve the goal of the research, the descriptive analysis method was used, and quantitative analysis was used. At the level of the descriptive analysis method, a desk survey was conducted. As for the quantitative analysis, it relied on applied data through a questionnaire form (Questioners) as a research tool, and the
... Show MoreBackground: Malignant lymphoma is the generic term given to tumors of the lymphoid system and specifically of lymphocytes and their precursor cells, While all lymphomas are malignant neoplasms, there is a wide spectrum of clinical behavior, with some following an indolent clinical course and others behaving in an aggressive manner (will causing death in a short time frame if left untreated). The metastatic process involves intravasation and extravasation of tumor cells, followed by reimplantation of tumor cells, formation of a new tumor stoma, Degradation of the extracellular matrix and components of the basement membrane by proteases facilitates the detachment of tumor cells, their crossing of tissue boundaries, and invasion into adjacent
... Show MoreIn this research, silver nanoparticles (AgNPs) were manufactured using aqueous extract of mushroom Pleurotus ostreatus. Anticancer potential of AgNPs was investigated versus human breast cancer cell line (MCF-7). Cytotoxic response was assessed by MTT assay. AgNPs showed inhibition effect at the following concentrations 12.5, 25, 50, 100 and 200 µg/ml versus MCF-7 cell line, and all treatments had a positive result. The MCF-7 cells were inhibited up to 85.14 % at the concentration 200 μg/ml of AgNPs which reduced cells viability to 14.86%, while 12.5 μg/ml of AgNPs caused 24.23% cells inhibition with reduction of cells viability to 75.77%.
Covalent modification of protein by drugs may disrupt self-tolerance, leading to lymphocyte activation. Until now, determination of the threshold required for this process has not been possible. Therefore, we performed quantitative mass spectrometric analyses to define the epitopes formed in tolerant and hypersensitive patients taking the β-lactam antibiotic piperacillin and the threshold required for T cell activation. A hydrolyzed piperacillin hapten was detected on four lysine residues of human serum albumin (HSA) isolated from tolerant patients. The level of modified Lys541 ranged from 2.6 to 4.8%. Analysis of plasma from hypersensitive patients revealed the same pattern and leve
A survey of chrysidid wasps and an identification key to the genera from different localities in Iraq are provided. The genus Pseudomalus Ashmead, 1902 with Pseudomalus auratus (Linnaeus) and Omalus timidus (Nurse), are new records from Iraq. As well, a preliminary checklist of Iraq’s Chrysididae is suggested. Twenty species within nine genera in two subfamilies have been included in the current list. Historical information is provided together with remarks on the current taxonomic status and the particular validity for some species.
Solar cells thin films were prepared using polyvinyl alcohol (PVA) as a thin film, with extract of natural pigment from local flower. A concentration of 0.1g/ml of polyvinyl alcohol solution in water was prepared for four samples, with various concentrations of plant pigment (0, 15, 25 and 50) % added to each of the four solutions separately for preparing (PVA with low concentrated dye , PVA with medium concentrated dye and PVA with high concentrated dye ) thin films respectively . Ultraviolet absorption regions were obtained by computerized UV-Visible (CECIL 2700). Optical properties including (absorbance, reflectance, absorption coefficient, energy gap and dielectric constant) via UV- Vis were tested, too. Fourier transform infra
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