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.
Interested current Research measuring damage currency Swap by converting The ministry of higher Education and scientific Research money The Iraqi dinar To U.S dollar by Trade Bank Of Iraq , And that The damage Generated resulting from Deferent Between the Exchange Rate adopted From Central Bank of Iraq and Market Exchange Rate adopted by The Trade Bank Of Iraq , and Which led to the greet damage ( losses ) in Bearing by the ministry, which led to the reduction of the financial allocations for licensed curriculum outside of Iraq , and this in turn leads to reduction in the number of students Sender ( scholarships ) outside Iraq.
Where the estimated loss (damage) that suffer by the Ministry of H
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In this study, the potential of adsorption of amoxicillin antibiotic (AMOX) from aqueous solutions using prepared activated carbon (AC) was studied. The used AC was prepared from an inexpensive and available precursor (sunflower seed hulls (SSH)) and activated by potassium hydroxide (KOH). The prepared AC was examined for its ability to remove AMOX from aqueous contaminated solutions and characterized with the aid of N2 -adsorption/desorption isotherm Brunauer–Emmett– Teller, scanning electron microscopy, energy-dispersive X-ray spectroscopy and Fourier-transform infrared. Zeta potential of the prepared activated carbon from sunflower seed hulls (SSHAC) were studied in relation to AMOX adsorption. The physical and chemical propert
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Abstract: Recombinant Newcastle disease virus (rNDV) has shown an anticancer effect in preclinical studies, but has never been tested in a lung cancer models. In this study we explored the anticancer activity of genetically modified NDV expressing IL-2-P53 (rClone30–IL-2-P53) in lung cancer model. We have cloned IL-2 and P53 genes and inserted them in the viral genome of New Castle Disease Virus to create a genetically modified rNDV- IL-2-P53 virus and tested the anti-tumor activity of the new virus in vitro on different types of cancer cell lines by MTT assay. TheIL-2 and P53 gene were successfully cloned and inserted into the viral genome by using a Mlu I and Sfi I endonucleases, viral vector was constructed correctly and successf
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Abstract
The current research is attempt to test the reflection of the lean management on the human resources management practices of two of the most important communication companies operating in Iraq (`Zain & Asia cell), The research aims to Determine the extent of adoption of the lean management approach in the two researched companies, as it improving human resource management practices. The research problem represented in the existence of lack of in some aspects of the application the lean management approach in service sector and neglecting the impact of its tools on the human resource management practices. For this purpose three principle research hypotheses has been formulated, first there is a correlation rel
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The current study introduces a novel technique to handle electrochemical localized corrosion in certain limited regions rather than applying comprehensive cathodic protection (CP) treatment. An impressed current cathodic protection cell (ICCPC) was fabricated and firmly installed on the middle of a steel structure surface to deter localized corrosion in fixed or mobile steel structures. The designed ICCPC comprises three essential parts: an anode, a cathode, and an artificial electrolyte. The latter was developed to mimic the function of the natural electrolyte in CP. A proportional-integrated-derivative (PID) controller was designed to stabilize this potential below the ICCPC at a cathodic potential of −850 mV, which is crucial for prote
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Photodynamic Action (PDA) by using appropriate wavelength of irradiation conjugated with porphyrin derivatives is a powerful mechanism of tumor destruction. Hematoporphyrin derivative has been shown to selectively localize in neoplastic cells and then cause destruction of them by generation of singlet oxygen when activated by low power He-Ne laser. Light which used in this study has been emitting from this laser has a wavelength equal to 632.8 nm (red light). Doses of laser had been varied from 3.6 J/cm2 to 14.4 J/cm2 . The beam of laser adjusted with a modified tissue culture plate. Cell lines had exposed to Hematoporphyrin D (HpD) for 24 hours before Laser exposure, their concentrations were varied from 5 µg/ml to 80 µg/ml. Resu
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New chelating ligand derived from triazole and its complexes with metal ions Rhodium, Platinum and Gold were synthesized. Through a copper (I)-catalyzed click reaction, the ligand produced 1,3-dipolar cycloaddition between 2,6-bis((prop-2-yn-1-yloxy) methyl) pyridine and 1-azidododecane. All structures of these new compounds were rigorously characterized in the solid state using spectroscopic techniques like: 1HNMR, 13CNMR, Uv-Vis, FTIR, metal and elemental analyses, magnetic susceptibility and conductivity measurements at room temperature, it was found that the ligand acts as a penta and tetradentate chelate through N3O2, N2O2, and the geometry of the new complexes are identified as octahedral for (Rh & Pt) complexes a
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