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.
Enterococcus faecalis is a natural inhabitant of the human gastrointestinal tract but can become dominant and cause infections when the intestinal homeostasis is disrupted. Enterococcal bacteria are considered one of the main reasons for the failure of endodontic treatment. This study aim to isolation and identification of E.faecalis depended on phenotype and molecular method, the phenotypic patterns using traditional biochemical methods, and then diagnosed it based on the genotypes and using specialized primers for 16srRNA and D-Ala: D-Ala ligase genes using polymerase chain reaction, In order to achieve successful treatment, it is necessary to study the bacterial behavior within the root canal system together with their resistance and def
... Show MoreThe way artists deal with body in their artistic works has had so many forms and methods, whether as an object for their drawings or as a material to create live artistic performances that relate to the idea of correspondence and interaction between different artistic categories such as: drama, dance, and painting as it is the case of the artist Marina Abramovic who has always used her body as an artistic unit to generate meaning and to perform her lively shows.
To go deeper into her career, our work was divided into 3 sections:
The first section was devoted to follow the main artistic stages that her body had gone through, starting with paintings she performed using concepts based on acting, simulation and nudity and ending wi
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Anaerobic digestion is a technology widely used for treatment of organic waste for biogas production as a source for clean energy. In this study, poultry house wastes (PHW) material was examined as a source for biogas production. The effects of inoculum addition, pretreatment of the substrate, and temperature on the biogas production were taken into full consideration. Results revealed that the effect of inoculum addition was more significant than the alkaline pretreatment of raw waste materials. The biogas recovery from inoculated waste materials exceeds its production from wastes without inoculation by approximately 70% at mesophilic conditions. Whereby, the increase of biogas recovery from pretreated wastes was by 20% higher than its
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The personality of the hero Salah al-Din al-Ayyubi (may God have mercy on him) came from the womb of jihad after difficult travails that the Arab Islamic nation experienced through the jihad of its loyal and honest sons who vowed themselves to God in defense of his religion and law, so between 490 AH - 540 AH outstanding jihadi leaders emerged who took upon themselves the responsibility of jihad and mobilizing the nation's energies To fight its enemies - the Franks, the Crusaders - in the Levant, and those leaders succeeded in achieving impressive victories over the Frankish military effort and regained some cities that were usurped by the Franks. Balak bin Bahram, Suqman, and Jakarmish, but these leaders could not maintain a state of un
... Show MoreParasitological examination of gills of three species of sparid fishes in the territorial waters of Iraq was performed, two diplectanid monogenoids were isolated and described; Lamellodiscus indicus Tripathi, 1959 from both Haffara seabream Rhabdosargus haffara (Forsskål, 1775) and Goldline seabream R. sarba (Forsskål, 1775) and Protolamellodiscus senilobatus Kritsky, Jiménez-Ruiz and Sey, 2000 from King soldierbream Argyrops spinifer (Forsskål, 1775). The record of the parasites is considered new to the parasite fauna of Iraq. The redescription of L. indicus for the first time which is collected from a new distribution area (Arabian Gulf). R. haffara is considered a new host record .
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