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 research study included the synthesis of a new series of heterocyclic derivatives containing the antibiotic Levofloxacin. The first way provides for the reaction of Levofloxacin with thionyl chloride in benzene as a solvent to give an acid chloride derivative. A new class of acid hydrazide synthesized from Levofloxacin was studied. Schiff bases were produced via the reaction of acid hydrazide with substituted aromatic ketones in methanol. The next stage involved the response of Schiff bases with thioglycolic acid and mono chloroacetic acid in DMF to produce derivatives of the antibiotic levofloxacin that have five heterocyclic members, including the derivatives thiazolidine-4-one and oxazolidine-5-one. The FTIR, 1HNMR, a
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Keys for 22 species representing ten genera Thripidae collection carried out during 1999-2001 in different localities in the middle of Iraq. Of them four species are described as new to science, Frankliniella megacephala sp. nov; Retithrips bagdadensis sp. nov; Chirothrips imperatus sp. nov; Taeniothrips tigridis sp. nov; Another thirteen species are recorded for the first time in Iraq; Thrips meridionalis (Pri.); Microcephalothrips abdominils (Crawford); Scolothrips pallidus (Beach); Scritothrips mangiferae Pri.; Frankliniella tritici Bagnall; Frankliniella schultzie Trybom; Frankliniella unicolor Morgan; Retithrips aegypticus Mar
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This research includes study of the effect of two kinds of Anthocyanin extracted , from extracted orange fruit ( Anthocyanin Evolvulus ,Methiola Violet ) on two types of pathological bacteria E.coli , staphylococcus aureus. The result shows that two kinds of extraction have nearly similar effect , and there is Inhibition zone of no growth between 10-12mm ,and the extraction (1) that has concentration of 10-3 mol./L is more effective..
Piroxicam is a non-steroidal anti-inflammatory drug (NSAID) used in the treatment of musculo-skeletal and joint disorders. The problem with this drug is its poor solubility in water and hence poor bioavailability after oral administration. In order to improve its solubility and dissolution behavior, hydrophilic additives such as starch, lactose, superdisintegrants including crospovidone (C.P), cross carmellose sodium (CCS), and sodium starch glycolate (SSG) were physically dry mixed with the drug by simple trituration. The improvement in the solubility in 0.1 N HCl was obtained as the amount of starch or lactose increased in the physical mixture, while for superdisintegrants, they further improve the solubility when they are present in s
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The research problem lies in identifying the challenges facing spreading the culture of peace in Iraq, and the most important factors of its instability from the point of view of leaders and intellectual elites.The research derives its importance from the fact that the peace imposed by justice leads to a stable Iraq and a prosperous future.
The study aims to identify the most prominent challenges and obstacles created by wars and faced the process of spreading the culture of peace and instability in Iraq, and to shed light on the reasons leading to the employment of the capabilities of community leaders in spreading the culture of peace.The objective of the current research is to identify the challenges of spreading the culture o
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The current research aims to identify the problems and needs for both college of political science and college of engineering’s students. The sample was (100) male and female student. The results showed bunch of problems which could be organized descendingly, the scientific domain ranged between (2 - 2.42), the mean of the psychological domain was (2.85), the moral domain ranged between (2.2 – 2.28)m the problems of study earned (2.30), the material domain got (1.95), the medical and social domain obtained (1.925), and finally, the family domain received (1.887).