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.
Increasing the power conversion efficiency (PCE) of silicon solar cells by improving their junction properties or minimizing light reflection losses remains a major challenge. Extensive studies were carried out in order to develop an effective antireflection coating for monocrystalline solar cells. Here we report on the preparation of a nanostructured cerium oxide thin film by pulsed laser deposition (PLD) as an antireflection coating for silicon solar cell. The structural, optical, and electrical properties of a cerium oxide nanostructure film are investigated as a function of the number of laser pulses. The X-ray diffraction results reveal that the deposited cerium oxide films are crystalline in nature and have a cubic fluorite. The field
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Objective: Zerumbone (ZER) is a well-known natural compound that has been reported to have anti-cancer effect. Thus, this study investigated the ZER potential to inhibit Thymidine Phosphorylase (TP) and the ability to trigger Reactive oxygen species (ROS)-mediated cytotoxicity in non-small cell lung cancer, NCI-H460, cell line. Material and Method: The antiangiogenic activity for ZER was evaluated by using the thymidine phosphorylase inhibitory test. Reactive oxygen species (ROS) production was determined via DCFDA dye by using flow cytometry. Result and Discussion: ZER was found to be potent TP inhibitory with the IC50 value of 50.3± 0.31 μg/ml or 230±1.42 µM. NCI-H460 cells upon treatment with ZER produced sign
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This research aims to study the effect of heat on the efficiency of solar cells of neutrons ranging from card to these cells in the case of dark and light before and after irradiation using the neutron source as well as electrical properties have been studied
Human interferon as is the case in all kinds of interferon has complex effects but all share their impact on preventing the proliferation of viruses and preventing or reducing human Alantervjørn conversion occurs if the cell is in preventing the growth of the virus when interferon Balnmstqubl connects
This study was conducted to investigate the effect of feeding diets containing different levels of parsley on the hematological traits of local Iraqi geese. A total of twenty-four local geese, one year old, were used in this experiment. The birds were allocated into four treatment groups, consisting of six geese each. Treatment groups were: control diet (C) (free from parsley); T1: control diet + 80 g/d parsley; T2: control diet + 160 g/d parsley; and T3: control diet + 240 g/d parsley. At the end of the experiment, blood samples were obtained from all geese from the brachial vein by venipuncture. Hematological traits included in this study were red blood cells count (RBC), hemoglobin concentration (Hb), packed cell volume (PCV), mean cell
... Show MoreIn this paper, we establish the conditions of the occurrence of the local bifurcations, such as saddle node, transcritical and pitchfork, of all equilibrium points of an eco-epidemiological model consisting of a prey-predator model with SI (susceptible-infected) epidemic diseases in prey population only and a refuge-stage structure in the predators. It is observed that there is a transcritical bifurcation near the axial and free predator equilibrium points, near disease-free equilibrium point is a saddle-node bifurcation and near positive (coexistence) equilibrium point is a saddle-node bifurcation, a transcritical bifurcation and a pitchfork bifurcation. Further investigations for Hopf bifurcation near coexistence equilibrium point
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This study was conducted at the poultry farm of the Department of Animal Production/College of Agriculture/University of Baghdad/Abu Ghraib, on 252 birds (180 females and 72 males). This study aims to observe the effect of melatonin implantation and exposure to different light colors and their interaction on characteristics of fertility and hatching of local Iraqi chickens. The birds were divided into three sections (white, red and green) each section contains two lines, one of which has been planted melatonin under the skin of the neck of birds and the other has not been planted hormones. The results showed that melatonin implantation and exposure to different light colors did not significantly affect the hatching rate of fertilized eggs a
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Background: Pain is one of the most postoperative complications of surgical wound especially within first 24 hrs. leading to delay hospital discharge, stress gastritis and increasing blood pressure. As wound infiltration with long acting local anesthetic (bupivacaine) has been proved to be effective after orthopedic surgeries especially total hip and knee replacements.Objective: our study was designed to determine theeffectiveness of local infiltration of 0.5% of bupivacainebefore closure of surgical wounds in controllingpostoperative pain and improve patient’s outcome after totalhip and knee replacement surgeries in first 24hrspostoperative period.Methods: Twenty patients from class I (healthy patients) and class II (patients mild sys
... Show MoreIn current study a computation fluid dynamic (CFD) technique was used to investigate the effect of groynes shape and spacing on the scour pattern and the maximum scour depth in open channel flow. CFD model have been validated throughout comparing the numerical results with three previous experimental studies for a single groyne located in open channel with three different shapes (L, quadrant, and parabola shapes). The comparison revealed very good agreement between numerical results of the maximum scour depth with the results of all experimental models. Moreover, investigations of the effect of multi-groynes (three groynes and four groynes) arranged in parallel with constant spacing and also with variable spacing have been done, the
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This experiment was conducted to study the effect of feeding diets containing different levels of parsley on the blood biochemical characteristics of local Iraqi geese. A total of twenty-four local geese, one year old, were used in this experiment during the period from the beginning of October to the end of December. The birds were allocated for Four treatment groups consisted of six geese each. Treatment groups were as follows: Control diet (T1) (free from parsley), T2: Control diet + 80 g / d parsley, T3: Control diet + 160 g / d parsley; T4: Control diet + 240 g / d parsley. At the end of experiment, blood samples were obtained from all geese by venipuncture from brachial vein and blood plasma samples were prepared. Blood biochemical tr
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