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 guava plant, Psidium guajava L., serves as proof of the abundant donations of nature, providing a delicious guava fruit; this plant is rich in groups of medicinal and nutritional benefits. Guava belonging to the Myrtaceae family, many previous studies reported many phytochemical constituents in its leaves that have many pharmacological activities and medicinal properties; this study focuses on the isolation, structural elucidation and calculation concentration of flavonoids, assessment of the cytotoxic activityof hyperin from Psidium guajava leaves newly cultivated in Iraq. The isolation process involved the use of thin-layer chromatography (TLC) and preparative high-performance liquid chromatography (PHPLC) and structural eluci
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Stevia rebaudiana Bertoni contains diterpenoid steviol glycosides that have no adverse impact on blood sugar levels despite being 300 times sweeter than sugar. This study aimed to investigate the rate of callus induction from stevia leaves and the content of glycosides when changing the sucrose percentage in the culture medium.. Murashige and Skoog (MS) culture medium supported by 4.0 mg/l naphthalene acetic acid (NAA) and 1.0 mg/l benzyl adenine (BA) was used, and different concentrations of sucrose (2, 3, 4, 5 and 6%) were tested .The extraction of glycosides from leaf and callus tissues was performed by using methanol. Extracted glycosides were analyzed by high-performance liquid chromatography (HPLC). The results showed signific
... Show MoreThe study aims to identify the reality of the use of educational technology in the teaching of mathematics from the point of view of mathematics teachers in Amman, where the researcher used the descriptive survey method. The sample included (67) female teachers working in public schools in Amman. In order to achieve the objectives of the study, the researcher used the questionnaire as a tool to collect data. The study concluded that many educational techniques are not available from the point of view of the study sample. Mathematics teachers also found that there are many obstacles that prevent the use of educational techniques in schools, most notably the lack of courses offered to teachers on the use of teaching techniques in teaching
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Some types of the fungus Aspergillus were isolated from some hospitals in the city of Baghdad (Imam Ali Hospital and Sadr General Hospital). The samples were taken by Transport media at a rate of three replicates of each place isolated from samples from different places within the hospital (waste, baths, the sick beds, corridors and room floors) for the purpose of isolating and diagnosing the fungus on the Czapeck Dox Agar media. It was noticed that the spread rate of fungus Aspergillus was 70% compared to other species that have emerged during the isolation process of the Sabouraud's Dextrose Agar media. The species A.niger (56.25%) was considered the most common type of fungus visible during the isolation process of the Imam Ali Hospit
... Show MoreThe research included four topics: the definition of Imam al-Bazzazi, his scientific life, his sheikhs, his students, his writings, his death, and selected issues of his weight in the book of prayer. The Imam al-Bazazi is the son of Muhammad ibnShihabibn Yusuf. He is known as al-Khardizi al-Khawarizmi. The author did not adopt the definitions of language and terminology, and rarely mention the legal evidence adopted by the scholars in their opinions, and only to mention the views only, and the author, may God have mercy on him in his book on the sources of m The Prophet (peace and blessings of Allaah be upon him) said: "The Prophet (peace and blessings of Allaah be upon him) And fall down the order, the third issue, which is to leave the
... Show MoreThis study aims to recognize the process of using computer in publicschools in Ma`an governorate in terms of hardware, capabilities, and teachers usage. It also aims to identify the most important obstacles those schools face from a teachers perspective.
The study sample consists of (150) teachers from different disciplines. The researcher prepared two questionnaires related to the research pivotal, then presented to a group of arbitrators to be used in this final form. The data was analyzed statistically and the study concluded that there is a shortage in the computer services provided to teachers, and there is a w
... Show MoreIn this work, new Schiff bases of quinazolinone derivatives (Q1-Q5) were synthesized from methyl anthranilate. The synthesis involved three steps. In the first step, methyl anthranilate was reacted with isothiocyanatobenzene, producing the thiourea derivative K1. The second step entailed reacting K1 with hydrazine hydrate, synthesizing 3-amino-2-(phenylamino) quinazolin-4(3H)-one (K2). The third step involved reaction of K2 with various aromatic aldehydes, yielding the Schiff bases derivatives Q1-Q5. The chemical structures of these compounds were identified by FT-IR,1H NMR and 13C NMR spectroscopy. The newly synthesized derivatives (Q1-Q5) were subjected to rigorous evaluation to assess their efficacy as corrosion inhibitors for ca
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The preparation of activated carbon (AC) from date stones by using microwave assisted K2CO3 activation was investigated in this paper. The influence of radiation time, radiation power, and impregnation ratio on the yield and methylene blue (MB) uptake of such carbon were studied. Based on Box-Wilson central composite design, two second order polynomial models were developed to correlate the process variables to the two responses. From the analysis of variance the significant variables on each response were identified. Optimum coditions of 8 min radiation time, 660 W radiation power and 1.5 g/g impregnation ratio gave 460.123 mg/g MB uptake and 19.99 % yield. The characteristics of the AC were examined by pore structure analysis, and scan
... Show MoreAbstract A total of 207 specimens were collected from different sources including patients, health care staff and hospital environment in Ibb city, Yemen. The study used the bacteriocin produced from active producer strains in typing of Staphylococcus aureus. Depending on the morphological, cultural and biochemical characteristics, 54 (26.09%) isolates of Staphylococcus aureus were identified. An antibiotic sensitivity test was done for the bacterial isolates, and the results showed that there were multiple resistant antibiotics. The Staphylococcin production of these isolates has been detected by using wells assay. Fifty one isolates were Staphylococcin producer. Four isolates (staph19, staph25, staph28 and staph43) were chosen as go
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