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.
This work investigates the effect of the gas nitriding process on the surface layer microstructure and mechanical properties for steel 37, tool steel X155CrVMo12-1 and stainless steel 316L. Nitriding was conducted at a temperature of 550 °C for 2 hours during the first stage and at 750 °C for 4 hours during the second stage. SEM and X-ray diffraction tests were performed to evaluate the microstructural features and the major phases formed after surface treatment. SEM and X-ray diffraction tests were performed to assess the microstructural features and the primary phases formed after surface treatment. The new secondary precipitates were identified as γ′-Fe4N, ε (Fe2–3N), and α-Fe, exhibiting an uneven chain-like pattern wit
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The present study provides the first record species of the genus Lithobius Leach, 1814, L. ferganensis (Trotzina, 1894) which was collected from the middle of Iraq. A detailed explanation of the morphology and the diagnostic characters of specimens of both sexes is provided.
The kinetic of atropine pertraction from seeds of Datura Metel Linn plant was studied. Diisopropyl ether, n-hexane and n-heptane were used as membranes for atropine recovery. The effect of speed of agitation and time in the range of 200-300 rpm and 0-3.5h, respectively were studied using the proposed membranes. The pertraction experiments were carried outs in a batch laboratory unit. The liquid-liquid pertraction was found to be very suitable for atropine recovery from its liquid extracts of Datura Metel seeds. A high purity (94-96%) can be obtained in the receiver phase. The pertraction process was found to be very selective for atropine recovery with diisopropyl ether membrane. As the speed of agitation increases the efficiency of pertrac
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Background: The possibility of converting the organic fraction of municipal solid waste to mature compost using the composting bin method was studied. Nine distinct treatments were created by combining municipal solid waste (MSW) with animal waste (3:1, 2:1), poultry manure (3:1, 2:1), mixed waste (2:1:1), agricultural waste (dry leaves), biocont (Trichoderm hazarium), and humic acid. Weekly monitoring of temperature, pH, EC, organic matter (OM percent), and the C/N ratio was performed, and macronutrients (N, P, K) were measured. Trace elements, including heavy metals (Cd and Pb), were tested in the first and final weeks of maturity. Results: Temperatures in the first days of composting reached the thermophilic phase in MSW compost
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Corncob is an agricultural biomass waste that was widely investigated as an adsorbent of contaminants after transforming it into activated carbon. In this research carbonization and chemical activation processes were achieved to synthesize corncob-activated carbon (CAC). Many pretreatment steps including crushing, grinding, and drying to obtain corncob powder were performed before the carbonization step. The carbonization of corncob powder has occurred in the absence of air at a temperature of 500 °C. The chemical activation was accomplished by using HCl as an acidic activation agent. Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), X-ray diffraction (XRD), and Brunauer–Emmett–Teller (BET) facilitate
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This study focused on the expression and regulation of BRCA1 in breast cancer cell lines compared to normal breast. BRCA1 transcript levels were assessed by real time quantitative polymerase chain reaction (RT-qPCR) in the cancer cell lines. Our data show overexpression of BRCA1 mRNA level in all the studied breast cancer cell lines: MCF-7, T47D, MDA-MB-231 and MDA-MB-468 along with Jurkat, leukemia T-lymphocyte, the positive control, relative to normal breast tissue. To investigate whether a positive or negative correlation exists between BRCA1 and the transcription factor E2F6, three different si-RNA specific for E2F6 were used to transfect the normal and cancerous breast cell lines. Interestingly, strong negative relationship was found b
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The research aimed to know favoured mass media for children and
modifying their behaviour ,the child became aquires the information from
mass media that he exposure them without any guidance , where upon the
quidance proqrammes becomes real danger whereas qet out their civil
style and converting to deadly poisons,and because of little study for this
supject the two researchers opined to perform astudy to know the favoured
mass media to the children and what are the mass media that modify their
behavior according to ther parent points of view ,after propring the research
measurement and the suilable statical methods it has shown that there are
mass media affect in children behavior ,they are st
This study aims to Statement of the relationship between Total Quality Management philosophy and Organizational performance from the point of view of the internal customer. A comparison has been made between two companies, one of which applies the requirements of TQM well and the other does not apply these requirements as the (General Company for Electrical Industries/ Diyala) and (General Company for Electrical Industries/ Baghdad) to conduct the search, During the questionnaire prepared for this purpose and distributed to a sample of 30 employees in the General Company for Electric Industries/ Diyala and (20) employees of the General Company for Electrical Industries/ Baghdad. Their answers were analyzed using a simple correlation coef
... Show MoreSome 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
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