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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The design of this paper is to find the possible correlation of Epstein Barr virus infection ina group of Iraqi women with cervical carcinoma though detection of Latent Membrane Protein 1 (LMP1) in these cervical tissues. Paraffinized blocks of two groups were included. The first sample of 30 cervical carcinomatous tissues and 15 biopsies from an apparently normal cervical tissues. All the samples were sectioned on a positive charged slides with 4 mm – thickness then submitted for immunohistochemical (IHC) staining to detect viral LMP1 expression. Sixty three percentage (19 out of 30) of the studies group showed positive overexpression as shown in with a significant association of the expression with cervical cancer with a significant ass
... Show Morehe study aimed to purify of Leucine aminopeptidase (LAP) from sera of hyperthyroidism patients and its relation to some thyroid hormones (TSH, T 3 , T 4 ) of subclinical hyperthyroidism and hyperthyroidism patients with lipid peroxidation levels that may be play a role in this diseases. Specimens were collected during the time from Nov 2017 until Jan 2018 from Endocrine and Diabetes Center, blood samples were collected from fifty healthy control and one hundred patients, patients were divided into two groups consisted of (50) with hyperthyroidism and (50) with subclinical hyperthyroidism. The aged for all subjects ranged (15-60) years with body mass ranged ((25- 29) kg/m 2 . The purification is done by addition of ammonium sulfate, dial
... Show MoreThis research aims to investigate and evaluate a reactive powder concrete (RPC) cast using economical materials. Its mechanical properties were investigated and evaluated by studying the effects of using different cement and silica fume contents and locally steel fibers aspect ratios as reinforcement for this concrete. A compressive strength of about 155.2MPa, indirect tensile strength of 16.0MPa, modulus of elasticity of 48.7GPa, flexural strength of 43.5MPa, impact energy of 3294.4kN.m and abrasion loss 0.59% have been achieved for reinforced RPC contains 910 kg/m3 cement content, silica fume content 185 kg/m3 of cement weight and fiber volume fraction 2%. The water absorption values w
... Show MoreThe study investigated the behaviour of asphalt concrete mixes for aggregate gradations, according to the Iraqi specification using the Bailey method designed by an Excel spreadsheet. In mixing aggregates with varying gradations (coarse and fine aggregate), The Bailey method is a systematic methodology that offers aggregate interlocking as the backbone of the framework and a controlled gradation to complete the blends. Six types of gradation are used according to the bailey method considered in this study. Two-course prepared Asphalt Concrete Wearing and Asphalt Concrete binder, the Nominal Maximum Aggregate Sizes (NMAS) of the mixtures are 19 and 12.5 mm, respectively. The total number of specimens was 240 for both layers (15 samp
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Conventional flexible pavements are released to different types of failure in the initial phases of their service life due to high traffic density, high speeds, heavy loads, and harsh climates. To eliminate pavement damage and failure early, the present search investigates the impact of adding glass, steel, and basalt fibers in the asphalt mixtures. Also, the study evaluates these materials characteristics compared to the mixtures without fibers. The Marshall test and tensile strength ratio test (TSR) were utilized to evaluate the asphalt mixture's performance. A set of specimens were produced by incorporating glass fiber (GF), steel fiber (SF), and basalt fiber (BF) at (0.10%, 0.15%, 0.20%), (0.25%, 0.35%, 0.45%), and (0.15%, 0.35%
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The isolates of Staphylococcus aureus were isolated from patients with various infections in hospitals, the isolates were identified and accurately diagnosed by phenotypic examination and biochemical tests, as well Vitek-2, and then genetic detection and diagnosis of many of the pathogenic factors associated with Staphylococcus aureus using conventional polymerase chain reaction (PCR) and testing for association by antibiotic resistance and production of some toxins by Staphylococcus aureus. After performing analysis of statistical, it was set up that the correlation coefficient of the PCR technique using virulence genes, sensitivity test to antibiotics and other virulence factors were significant at p < 0.05, but was insignificant with the
... Show MoreSteps were taken to obtain the Kojic acid crystals from local fungal isolation A. flavus WJF81 by separating the fermentation products from the fungus mycelium from the production plant at the centrifuge at a speed of 5000 cycles for 10 minutes. The extraction was followed by ethyl acetate then supernatant concentrate by using rotary evaporator, and dried with heat oven 37ºC. Long, yellowish, pristine acid crystals were obtained that examined the optical microscope with a magnification force of 10x and 40x. The melting point of kojic acid was determined between 152.9-153.5 °C Results of the diagnosis of Kojic acid by applying High pressure liquid chromatography HPLC technique showed that the acid was at one peak, which was close to the
... Show MoreIn the present work is the deposition of copper oxide using the pulsed laser deposition technique using Reactive Pulsed Laser as a Deposition technique (RPLD), 1.064μm, 7 nsec Q-switch Nd-YAG laser with 400 mJ/cm2 laser energy’s has been used to ablated high purity cupper target and deposited on the porous silicon substrates recorded and study the effect of rapid thermal annealing on the structural characteristics, morphological, electrical characteristics and properties of the solar cell. Results of AFM likelihood of improved absorption, thereby reducing the reflection compared with crystalline silicon surface. The results showed the characteristics of the solar cell and a clear improvement in the efficiency of the solar cell in the
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