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.
Advancements and modernizations introduced into the educational and pedagogical systems have significantly impacted teaching processes and how subjects are presented and explained to students. The focus has shifted to how learners interact with the material they need to learn, providing sufficient opportunities for learning and granting them freedom and self-confidence to achieve learning objectives. The research problem stems from the researcher's experience as a lecturer in the College of Physical Education and Sports Science, particularly in teaching basketball. She observed that some instructors were deficient in using the most effective teaching methods. The researcher formulated her research question based on these observations: "What
... Show MoreRecently, there has been an increase in the prevalence of ulcerative colitis (UC), and inflammatory bowel diseases (IBD) worldwide, especially in certain recently industrialized countries like China and In¬dia. Globally, the prevalence of UC, a chronic illness that affects the large intestine, is rising. Fifty adherent invasive Escherichia coli (AIEC) isolates were identified from ulcerative colitis biopsy samples originating from the Gastrointestinal tract (GIT) and Hepatology teaching hospitals/medical city in Baghdad City. The test’s results demonstrated that the AIEC isolates had a high level of resistance to the majority of the an-tibiotics under investigation. Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR) and m
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Background: Most primary Health Care Centers (PHCCs) in Iraq have a referral system records; however, this mechanism does not function well because of the lack of other requirements for an efficient referral system.
Objective: To assess the practice & opinion of doctors in PHCs toward the referral system, and to determine the doctors in PHC's commitment to referral system instructions and guidelines.
Subjects and methods: A cross-sectional study with analytic elements was conducted in nine health directorates in Iraq, from the 1st October 2018 – 30th June 2019.One PHC was selected randomly form each sector in every governorate, A questionnaire was used to collect the required information.
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تم استخدام خرائط ضبط الجودة الإحصائية لتقييم جودة الخدمة التعليمية في جامعة الباحة، ويهدف هذا البحث إلى استخدام خرائط ضبط الجودة الإحصائية لقياس مستوى الجودة وفجوة الجودة بين توقعات الطلبة وإدراكاتهم لمستوى الخدمة الذي تقدمه جامعة الباحة. حيث تم اختيار عينة من 200 طالب وطالبة عشوائيا باستخدام العشوائية العنقودية من 4 كليات خلال الفترة 01 – 30/2015م، وجمعت البيانات من خلال استبيان جودة الخدمة الذي يقيس ت
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This experiment presented essential oils by GC/MS, pigment content, and their antioxidant activities as well as sensory evaluation of delight samples. Limonene (66.88%) was the most prevalent yield. The peels of clementine had DPPH and ABT Scavenging activity. All levels of pigment extract had better scores for all sensory values and recorded acceptable scores in terms of appearance, color, aroma, and overall acceptability compared to control delight. Besides, delight samples containing 15 mg astaxanthin pigment extract showed maximum sensory scores compared to other samples and control delight. On the other hand, the product was less acceptable to the panelists compared to control in the case of the addition of 3.75 mg astaxanthin pigme
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Release of industrial effluents comprising dyes in water bodies is one of the foremost causes of water pollution. Therefore, the proper and proficient treatment of these dyes contaminated left-over material before their release is crucial. Herein, an eco-friendly biological macromolecule Gum-Acacia (GA) integrated Fe3O4 nanoparticles composite hydrogel was manufactured via co-precipitation technique for effective adsorption of Congo red (CR) dye existing in water bodies. The as-prepared magnetic GA/Fe3O4 composite hydrogel was characterized by FTIR, XRD, EDX, VSM, SEM, and BET techniques. These studies discovered the fruitful fabrication of biodegradable magnetic GA/Fe3O4 composite hydrogel possessing porous structure with large surface are
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