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.
Euphemisms are advantageous in people’s social life by turning sensitive into a more acceptable ones so that resentful feelings and embarrassment can be avoided. This study investigates the ability of Iraqi English learners in using euphemistic expressions, meanwhile, raising their awareness and the faculty members in English teaching faculties regarding the relevance of discussing the topics that demand euphemisation. This study comprised three stages: initial test, explicit instruction with activities, and a final test for the students’ development in this domain. A test has been distributed among 50 respondents, who are at the fourth year of their undergraduate study at the University of Babylon/ College of Basic Education. The lo
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Uropathogenic Escherichia coli is the main cause of urinary tract infections, the ability of this bacteria to cause urinary tract infections is related to a variety of virulence factors that enhance colonization and evade the immune response, one of these virulence factors is cytotoxic necrotizing factor 1 toxin which converts the glutamine residue to glutamic acid to activated GTPase Rho family. The study was meant to find out the prevalence rate of the cnf1 gene in Uropathogenic Escherichia coli isolated from Iraqi patients. Conventional laboratory methods were used for primary bacterial identification and molecular methods were used to confirm bacterial identity and gene detection. Escherichia coli was identified in 89/165 (53.93%) of th
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Three types of extracts ( aquatic, alcoholic, and oily ) were prepared from the fruits of coconuts, and a series of chemical tests were conducted in addition to the use of the FTIR equipment to determine the active locations in the prepared extracts. The results indicated the presence of active compounds (tannins, saponins, flavonoids, turbines and steroids) in the extracts prepared from the fruits of coconuts, also the antimicrobial capability of these extracts were tested on pathogenic bacteria isolated from wounds and burns infections cases. The results proved that the concentration 80 mg/ml of the aquatic extract is the minimum inhibitory concentration for the microbes: Proteus vulgaris and Pseudomonas fluorescence, while the
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A common field development task is the object of the present research by specifying the best location of new horizontal re-entry wells within AB unit of South Rumaila Oil Field. One of the key parameters in the success of a new well is the well location in the reservoir, especially when there are several wells are planned to be drilled from the existing wells. This paper demonstrates an application of neural network with reservoir simulation technique as decision tool. A fully trained predictive artificial feed forward neural network (FFNNW) with efficient selection of horizontal re-entry wells location in AB unit has been carried out with maintaining a reasonable accuracy. Sets of available input data were collected from the exploited g
... Show MoreStaphylococcus lugdunensis, isolation between 12.5 to 1.8% routine works may be a possible peroral route of infective endocarditis and found in the oral cavity by examined using saliva. Similar supragingival plaque isolation was observed. The increased bacteria resistance to antibiotics multiple have led to novel methods for resistance bacteria; antimicrobial agents are well known (ZnO NPs) by biological method and are lower toxicity and biology safety ZnNOPs activity by plant extraction and less toxicity as well as bio-safe. The nanoparticle was synthesized by biological method (Green) by barberry (Berberis vulgaris) extract. In this study using (WAD) method using different concentrations between (128, 64, 32, and 16) mg/mL of ZnO NPs, The
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Staphylococcus lugdunensis, isolation between 12.5 to 1.8% routine works may be a possible peroral route of infective endocarditis and found in the oral cavity by examined using saliva. Similar supragingival plaque isolation was observed. The increased bacteria resistance to antibiotics multiple have led to novel methods for resistance bacteria; antimicrobial agents are well known (ZnO NPs) by biological method and are lower toxicity and biology safety ZnNOPs activity by plant extraction and less toxicity as well as bio-safe. The nanoparticle was synthesized by biological method (Green) by barberry (Berberis vulgaris) extract. In this study using (WAD) method using different concentrations between (128, 64, 32, and 16) mg/mL of ZnO
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This research examines how Al-Sabah newspaper addressed violations of women’s rights in Iraq. Using content analysis, a sample of 157 articles published from July 1, 2021, to December 31, 2021, was analyzed. The study reveals diverse journalistic treatments, with interpretive approaches employed in addressing violations of Iraqi women’s rights. Social violations of women’s rights were the primary focus, and various forms of newspaper articles, including opinion pieces, were commonly used.