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.
A synthesis series of new heterocyclic derivatives (A2-A7) (pyrrole, pyridazine, oxazine and imidazol) derived from 4-acetyl-2,5-dichloro-1-(3,5-dinitrophenyl)-1H-pyrrole-3-carboxylate(A1) have been synthesised. Synthesis of compound (A2) by the reaction of starting material (A1) with hydroxyl amine hydrochloride in the presence of pyridine. Compound (A2) was reacted with hydrazine hydrate in dry benzene to give (A3) derivative. The compound )A3( deals with sodium nitrite to give diazonium salt, and the reaction diazonium salt with ethyl acetoacetate to produce compound (A4). To a mixture of compound (A4) and hydroxyl amine with sttired to yield (A5).Compound (A6) was prepared by reaction compound (A4) with thiosemicarbazide in presence
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Klebsiella pneumoniae has been found in the urinary tract of some bladder cancer patients. Bacterial presence within tumor tissue may affect the tumor-microenvironment and consequently influence cancer behavior, development, and treatment response. This study investigated mesenchymal and stemness transdifferentiation of bladder cancer cell line due to environmental stress of K. pneumoniae. Cultures of urothelial bladder cancer cell line (T24) were infected with K. pneumoniae with different multiplicity of infection (MOI) for two and four days. Transdifferentiation-associated features were morphologically assessed.
Moreover, transdifferentiation markers were estimated using Q-PCR and immunohistoc
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total of 17 Escherichia coli isolates were collected from urine specimens of patients with urinary tract infection. Antibiotics sensitivity test indicated that amikacin followed by chloramphenicol and ciprofloxacin are the most effective antibiotics. The isolates showed multidruge resistant, nine isolates were resistant to 11-15 antibiotics, 3 were resistant to 16-20 antibiotics and 5 were resistant to 21-25 antibiotic. Two isolates were selected, the first (ED1) was resistant to (22) antibiotics while the second isolate (ED2) was resistant to (14) antibiotics (out of 25). Minimum inhibitory concentration (MIC) of the black and green tea water boiled extracts were determined towards (ED1,ED2).Results showed that MIC of black tea extr
... Show MoreThis effort is related to describe and assess the performance of the Iraqi cement sample planned for oil well-cementing jobs in Iraq. In this paper, major cementing properties which are thickening time, compressive strength, and free water in addition to the rheological properties and filtration of cement slurry underneath definite circumstances are experimentally tested. The consequences point to that the Iraqi cement after special additives encounter the requests of the API standards and can consequently is used in cementing jobs for oil wells. At this research, there is a comparative investigation established on experimental work on the effectiveness of some additives that considered as waste materials which are silica fume, baux
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This research is studying technique sculptures super - realism, search through, how the method of work, and the search for the materials used in their manufacture, and this is the first study in the field of art and the field of academic study in the country.Research consists of an introduction, And four sections, The introduction containing information on: research problem, Importance of research, Goals of the research, Limits of research, research approach, and research tools.The first section contains a technical study sculptures super -realism in contemporary sculpture, while the second section includes a search for alternative materials available in the local markets, for making sculptures super - realism, the third section dedicate
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This effort is related to describe and assess the performance of the Iraqi cement sample planned for oil well-cementing jobs in Iraq. In this paper, major cementing properties which are thickening time, compressive strength, and free water in addition to the rheological properties and filtration of cement slurry underneath definite circumstances are experimentally tested. The consequences point to that the Iraqi cement after special additives encounter the requests of the API standards and can consequently is used in cementing jobs for oil wells. At this research, there is a comparative investigation established on experimental work on the effectiveness of some additives that considered as waste materials which are silica fume, bauxite,
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Gaining the relationship between the central government and local governments is great importance especially after the expanded functions of the central government and became unable to carry out their works without all that delegate or transfer parts of its powers to lower levels.
The aims of the research is to determine the relationship of the central government with the local government in accordance with the principle of administrative decentralization with the presentation of the competent authorities, the control of local governments, according to the Provincial Council Act No. 21 of 2008 amended.
The research has a problem that has been the relationship of the central
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Dust samples have been collected from three areas in Baghdad during dust storm occurred in 18th of June 2009 to characterize elemental particle size and composition by different techniques. The x-ray diffraction detected six minerals those are calcite, and quartz, present as a major components, dolomite, kaolinite, gypsum and plagioclase present as miner components .EDX detected some normal elements presented in local soil except traces of lead , nickel, and chromium. The particle size analysis by a set of sieves have revealed that the majority particle distribution was between (32 and 45)μm . To isolate the aerosol size, PM10 buoyancy method of powder in water showed a signifying amounts of particulate size .Scheerer’s method was app
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