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 - M ultiple mixing ratios of -transitions from levels of 56Fe populated in 56 56 Fe n n Fe ( , ) reactions are calculated by using const. S.T.M. This method has been used in other works [3,7] but with pure transition or with transitions that can be considered as pure transitions، in our work we used This method for mixed - transitions in addition to pure - transitions. The experimental angular distribution coefficients a2 was used from previous works [1] in order to calculet - values. It is clear from the results that the - values are in good agreement or consistent, within associated errors, with those reported previously [1]. The discrepancies that occur are due to inaccuracies existing in the expe
... Show MoreIn the present work, the phthalic acid (phthH2) and 1.10 phenonthroline (phen), and their complexes were synthesized and isolated as [M(phth)(phen)2], Mn(II), Fe(II), Co(II), Ni(II) Cu(II), Zn(II), and Cd(II) ions. These complexes were characterized by elemental analysis, melting point, conductivity, percentage metal, UV–Vis, FT-IR, and magnetic moment measurements. The molar conductance indicates that all the metal complexes in DMSO are nonelectrolytic. phthalic acid (phtha), and 1,10-Phenanthroline (phen), behaved as bidentate, coordinating to the metal ion through their two oxygen and two pyridinyl nitrogen atoms respectively, as corroborated by. Electronic spectra, FTIR, spectroscopy amusement indicated that all the metal complexes ad
... Show MoreThe present work reports on the performance of three types of nanofiltration membranes in the removal of highly polluting and toxic lead (Pb2+) and cadmium (Cd2+) from single and binary salt aqueous solutions simulating real wastewaters. The effect of the operating variables (pH (5.5-6.5), types of NF membrane and initial ions concentration (10-250 ppm)) on the separation process and water flux was investigated. It was observed that the rejection efficiency increased with increasing pH of solution and decreasing the initial metal ions concentrations. While the flux decreased with increasing pH of solution and increasing initial metal ions concentrations. The maximum rejection of lead and cadmium ion
... Show MoreBackground: A diverse group of bacteria live in biofilms in the oral cavity. On dental surfaces biofilms form plaque that is potentially involved in caries and periodontal diseases. Periodic studying of plaque microflora and their antimicrobial sensitivity patterns strongly affects the clinical practice in plaque-induced oral diseases. Materials and methods: Dental plaque samples were collected from 22 patients having ages ranged between 33 and 49 years with gingivitis that met the study criteria. Plaque, gingival and gingival bleeding indices (PI, GI, GBI) were measured for each patient. Laboratory procedures included microbiological examination of plaque samples followed by antibiotic sensitivity testing using disc diffusion method were
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Bacterial strains were isolated from oil-contaminated soil, in 2018, these isolates were identified, and with the aim of finding out the ability of these isolates to degrede the oil compounds, the color change of medium which added to it isolates was read by the method of Pacto Bushnell Hans. Then the change in the petroleum compounds was read by gas chromatography, for the most effective isolates.
The nine isolated bacterial showed different degrees of color change, and the isolates (Pseudomonas, Bacillus, Micrococcus) outperformed the color change amount (78, 78, 77) %, respectively, compared to the control, and the three isolates together showed the best color change of 90.7. % Compared to the control, and the
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The remove of direct blue (DB71) anionic dye on flint clay in aqueous solution was investigated by using a batch system for various dye concentrations. The contact time, pH, adsorbent dose, and temperature was studied under batch adsorption technique. The data of adsorption equilibrium fit with isotherm Langmuar and Freiundlich ,when the correlation coefficient used to elucidate the best fitting isotherm model. The thermodynamic parameters such as, ?Hº ,?Sº and ?Gº. Thermodynamic analysis indicated that the sorption of the dyes onto Flint clay was endothermic and spontaneous.
Background: Esthetic treatment is the options of patient seeking orthodontic treatment. Therefore this study was conducted to measure the concentration of Aluminum, Nickel, Chromium and Iron ions released from combination of monocrysralline brackets with different arch wires immersed in artificial saliva at different duration, to evaluate the corrosion point on different parts of the orthodontic appliances before and after immersion in artificial saliva, and to evaluate the corrosion potential of each group of the orthodontic appliances. Material and methods: Eighty orthodontic sets prepared. Each set represents half fixed orthodontic appliance, from the central incisor to the first molar, for the maxillary arch, each set consisted of molar
... Show MoreA new 4-thiazolidinone, substitutedbenzylidene-thiazolidinone and tetrazole were synthesized from thiosemicarbazone and hydrazone. The thiosemicarbazone was prepared by the reaction of thiosemicarbazide with aldehyde derivative from L-ascorbic acid in absolute ethanol using glacial acetic acid as a catalyst. 1, 3-thiazolidin-4-ones were synthesized from the condensation of thiosemicarbazones with chloroacetic acid in presence of anhydrous sodium acetate. A 1, 3- thiazolidine-4-one was reaction with several 4-substitutedaldehydes to produce new derivatives with a double bond at the position-5 of the 4-thiazolidinone ring. While the tetrazole compounds were synthesized by 1, 3-cycloaddition reaction of sodium azide and hydrazone compounds in
... Show MoreFrom the sustainability point of view a combination of using water absorption polymer balls in concrete mix produce from Portland limestone cement (IL) is worth to be perceived. Compressive strength and drying shrinkage behavior for the mixes of concrete prepared by Ordinary Portland Cement (O.P.C) and Portland limestone cement (IL) were investigated in this research. Water absorbent polymer balls (WAPB) are innovative module in producing building materials due to the internal curing which eliminates autogenous shrinkage, enhances the strength at early age, improve the durability, give higher compressive strength at early age, and reduce the effect of insufficient external curing. Polymer balls (WAPB) had been used in the mixes of thi
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