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.
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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In this paper, the concept of contraction mapping on a -metric space is extended with a consideration on local contraction. As a result, two fixed point theorems were proved for contraction on a closed ball in a complete -metric space.
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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.
The research aimed to identify the effectiveness of instructional design according to whole brain theory of Herman in the achievement of chemistry at the fifth scientific students at a secondary school of the General Directorate for Educational in Diyala / Baladruz in Iraq. The research sample Consisted of (57 student, (29) students as experimental group studied according to instructional design strategies for whole brain theory of Herrmann and (28) a student as a control group studied by the usual way for two semesters, a prepared achievement test as article and objective type of multiple choice, the coefficient stability of alpha-Cronbach equation reached (0.86). The research Results showed the presence of a statistically significant d
... Show MoreFrom 211 urine samples, Gram negative bacteria were isolated from only 61 urine samples with isolation percentage 28.9%. Escherichia coli were isolated percentage 70.49% while Klebsiella pneumoniae and Psendomonas aeruginosa were 8.19% and 6.55%, respectively.Proteus spp. Were isolated from 9 (14.75%), P. mirablis and P. vulgaris were isolates percentage 11.47% and 3.27%, respectively. Uroepithelial Cell Adhesin (UCA) fimbriae expression by P.mirabilis isolates was detected by the high capacity to adhesion to human uroepithetial cells, the isolate p.mirabilis U7 was adhesion to human uroepithelial cells mean no.30.2 bacteria/cell when grown on luria broth at 37C for 24h, but then grown it’s on luria agar at 37C for 24h the adhesion
... Show MoreProtein bound fucose (PBF), protein bound hexose (PBHex), and total calcium {T.Ca) were 'determined in sera of (40) hy-p.ertnyroidism , (40) hypothyroidism patients and (40) controL The resultsr vealed a significant decrease in the kwel of PBF, PBHex and T.Ca in sera of patients with .hyperthyroidism compared to control; Inc se Qf PBF. there nQ difference in its level betwe.en patients with hypothyroidism and control group. While there is a significant increment in PBHex leveli:n both hyper and hypothyroidism With respect to that of control Result indicates, that total calcium levels were i.n the nomml range for all p tients groups. Patient compa
... Show MoreIntroduction: The study was intended for Roseomonas gilardii NTCC 13290 strain pigment extraction and characterization. Methodology: The pigment-producing bacterial were cultured on Columbia blood agar and nutrient media agar. Then the pigments were extracted by ethanol. The candidate pigment was further characterized by different biotechnological techniques: UV-Vis spectroscopy, FT-IR to analyze the functional group of the targeted pigment, and TLC media. Results: The cultivation of Roseomonas gilardii on media showed pink color and nearly runny texture. The bacterial colonies were microscopically gram stained and examined, the R. gilardii was seen as coccobacillus colonies that mostly form pairs arranged as short chains. The R. gilardii b
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Abstract. Silver, Indium Selenium thin film with a thickness (5001±30) nm, deposited by thermal evaporation methods at RT and annealing3temperature (Ta=400, 500 and 600) K on a substrate of glass to study structural and optical properties of thin films and on p-Si wafer to fabricate the AgInSe2/p-Si heterojunction solar cell. XRD analysis shows that the AgInSe2 (AIS) deposited film at RT and annealing3temperature (Ta=400, 500 and 600) K have polycrystalline structure. The average grain size has been estimated from AFM images. The energy gap was estimated from the optical transmittance using a spectrometer type (UV.-Visible 1800 spectra photometer). From I-V characterization , the photovoltaic parameters such as, open-circuit voltage, short
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In this paper, we consider a new approach to solve type of partial differential equation by using coupled Laplace transformation with decomposition method to find the exact solution for non–linear non–homogenous equation with initial conditions. The reliability for suggested approach illustrated by solving model equations such as second order linear and nonlinear Klein–Gordon equation. The application results show the efficiency and ability for suggested approach.