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.
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Background: Non-small cell lung cancer (NSCLC) is caused of 85% of all lung cancers. Among the most important factors for lung tumor growth and proliferation are the tyrosine kinase receptors that coded by the epidermal growth factor recep-tor (EGFR) gene. Activation of EGFR ultimately leads to developing of lung cancer. The present study was undertaken with an objective to detect EGFR mutations in bronchial wash from Iraqi patients with NSCLC before treatment. Methods: DNA was extracted from bronchial wash samples collected from 50 patients with NSCLC by using a Qiamp DNA Mini Kit (Qiagen, Hilden, Germany). Then, EGFR mutations were determined by using real-time RCR combined with two technologies, Amplification Refractory Mutation System (
... Show MoreBackground: Non-small cell lung cancer (NSCLC) is caused of 85% of all lung cancers. Among the most important factors for lung tumor growth and proliferation are the tyrosine kinase receptors that coded by the epidermal growth factor recep-tor (EGFR) gene. Activation of EGFR ultimately leads to developing of lung cancer. The present study was undertaken with an objective to detect EGFR mutations in bronchial wash from Iraqi patients with NSCLC before treatment. Methods: DNA was extracted from bronchial wash samples collected from 50 patients with NSCLC by using a Qiamp DNA Mini Kit (Qiagen, Hilden, Germany). Then, EGFR mutations were determined by using real-time RCR combined with two technologies, Amplification Refractory Mutation System (
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The structural, optical and photoelectrical properties of fabricated diffusion heterojunction (HJ) solar cell, from n-type c-Si wafer of [400] direction with Boron, has been studied. AgAl alloys was used because of its properties that affect as a good connection materials. TiO2 has been used as a reflecting layer to increase the absorption radiation. The HJ has direct allowed energy gap equal to 3.1 eV. The c-Si/B HJ solar cell yielded has an active area conversion efficiency of 16.4% with an open circuit voltage of (Voc) 0.592V, short circuit current (Isc) of 2.042mA, fill factor (F.F) of 0.682 and % =10.54.
In this work, strains and dynamic crack growth were studied and analyzed in thin flat plate with a surface crack at the center, subjected to cycling low velocity impact loading for two types of aluminum plates (2024, 6061). Experimental and numerical methods were implemented to achieve this research. Numerical analysis using program (ANSYS11-APDL) based on finite element method used to analysis the strains with respect to time at crack tip and then find the velocity of the crack growth under cycling impact loading. In the experimental work, a rig was designed and manufactured to applying the cycling impact loading on the cracked specimens. The grid points was screened in front of the crack tip to measure the elastic-plas
... Show MoreBy using governing differential equation and the Rayleigh-Ritz method of minimizing the total potential energy of a thermoelastic structural system of isotropic thermoelastic thin plates, thermal buckling equations were established for rectangular plate with different fixing edge conditions and with different aspect ratio. The strain energy stored in a plate element due to bending, mid-plane thermal force and thermal bending was obtained. Three types of thermal distribution have been considered these are: uniform temperature, linear distribution and non-linear thermal distribution across thickness. It is observed that the buckling strength enhanced considerably by additional clamping of edges. Also, the thermal buckling temperatures and
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The study aimed to increase the biological value of white bean. The effect of different concentrations 0.01 ,0.02,0.03,and 0.04 M of sodium sulfite solutions for 1hr at 70 ºC on the trypsin inhibitors activity, protein isolate and protein solubility of complete and dehulling white bean flour were studied.Trypsin inhibitors activity were reduced by 42.97, 58.69, 68.59 and 69.58% in complete white bean flour at 0.01 ,0.02, 0.03, 0.04 M respectively, while the corresponding values were 50.43, 61.00, 75.61 and 85.66% respectively in dehulling white bean flour .Protein isolate value was 13.41% and protein solubility was 2.2% in control sample, Furthermore, the using of chemical treatment showed that protein isolate was reduced gradually and
... Show MoreSurface water flow samples were collected with distances downstream over Saqlawiya main drain whose stretch of about 24.5 km. The drain travels through different land use pattern, before, flowing into Tigris River. Eight sampling points were carefully
selected downstream the channel during dry season. The examined water parameters were pH, NH3, NO3, PO= 4, BOD5, COD, TDS, S.S, Cl-, SO= 4, Na+ , Ca+2, Mg+2, and Oil and Grease. Descriptive and inferential methods through finding the best curve fit correlation were employed in the study to test the strength of the association between water chemical characteristics and distance downstream the channel. A comparison of the values of chemical parameters at the Al-Saqlawiya Drain-Tigris Riv