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.
Nowadays, most of the on-chip plasmonic single-photon sources emit an unpolarized stream of single photons that demand a subsequent polarizer stage in a practical quantum cryptography system. In this paper, we numerically demonstrated the coupling of the light emitted from a quantum emitter (QE) at 700 nm wavelength to the propagation mode supported by an on-chip hybrid plasmonic waveguide (HPW) polarization rotator. Our results proved that the light emitted is linearly polarized at 0º, 45º/−45º, and 90º with propagation lengths of 5 μm, 3.3 μm, and 3.9 μm, respectively. Moreover, high power-conversion efficiency was obtained from an applied transverse magnetic (TM) mode (0º-polarization) to a transverse electric (TE) (90º-polari
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A simple, rapid and environmentally friendly dispersive liquid–liquid microextraction method-based spectrophotometric method for the trace determination of folic acid has been developed. The proposed method is based on the formation of a deep yellow product via reaction of folic acid and 1,2-naphthoquine-4-sulfonate at pH = 9. The formed complex was extracted using a mixture of chloroform and ethanol. Then, the tiny organic droplets were measured at λ = 520 nm. At the optimum conditions, linearity was ranged from 0.05 to 1.5 μg/mL for the standard and samples, with a linear correlation coefficient of 0.9996. The detection limits were 0.02, 0.027, 0.03, 0.02 and 0.04 μg/mL for standard, tablet (5 mg), tablet (1 mg), syrup and fl
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The wastewater arising from pulp and paper mills is highly polluted and has to be treated before discharged into rivers. Coagulation-flocculation process using natural polymers has grown rapidly in wastewater treatment. In this work, the performance of alum and Polyaluminum Chloride (PACl) when used alone and when coupled with Fenugreek mucilage on the treatment of pulp and paper mill wastewater were studied. The experiments were carried out in jar tests with alum, PACl and Fenugreek mucilage dosages range of 50-2000 mg/L, rapid mixing at 200 rpm for 2 min, followed by slow mixing at 40 rpm for 15 min and settling time of 30 min. The effectiveness of Fenugreek mucilage was measured by the reduction of turbidity and Chemical Oxygen Demand
... Show MoreThis study involves the investigation of the effect of nitrogen laser with 337.1 nm wavelength on the sensitivity of Staphylococcus aureus bacteria by using local therapeutic due to burns. Thirty six isolate of Staphylococcus aureus bacteria were isolated from 25 patients suffering from sever burns, each isolate of bacteria was irradiated with nitrogen laser at (5, 10, 15 and 30) pulses/second repetition rates for 1, 5, 10, 20 and 30 minutes for each repetition rate. The effects of nitrogen laser on the local therapeutics sensitivity of bacteria were obtained using Kirby Baur method. Changes in the sensitivity of bacteria to local therapeutics (Tetracyclin, Chloramphenicol, Flumizin and Fucidin) occur at high repetition rate(30 pulses/seco
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Prodigiosin is a ‘natural red pigment produced by Serratia marcescens which exhibits immunosuppressive and anticancer properties in addition to antimicrobial activities. This work presents an attempt to maximize the production of prodigiosin by two different strategies: one factor at time (OFAT) and statistical optimization. The result of OFAT revealed that sucrose and peptone were the best carbon and nitrogen sources for pigment production with concentration of prodigiosin of about 135 mg/ L. This value was increased to 331.6mg/ L with an optimized ratio of C/N (60:40) and reached 356.8 with pH 6 and 2% inoculum size at end of classical optimization. Statistical experimental design based on Response surface methodology was co
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?Natural Occurrence of aflatoxin Bi and B2 was investigated monthly in paddy rice stored for duration of six months. Aflatoxin B| was detected at the beginning of storage. The concentration of aflatoxin Bi in paddy and polished rice was found to be (0.3 and ()) ppb respectively while aflatoxin B2 W'as not detected in both paddy and polished rice afler six months of storage, concentration of aflatoxin Bi ‘ ’ ' reached 3 ppb in paddy rice and 0.9 ppb in polished rice, while the study proved that the concentration of aflatoxin in paddy rice was higher than polished rice as a result of processing which reduce the toxin
This dissertation explores the role of Iraqi E-press in crystallizing the orientations or directions of the public towards the local political issues like (demonstrations, parliamentarian elections, provincial elections, the public budget and its consequences, Iraq’s relations with neighboring countries, Iraqi HOR and its sessions, and the relation of the federal government with KRG). The dissertation’s main problem revolves around a central inquiry: what is the role of the Iraqi E-press in crystalizing the direction of the Iraqi public towards the local political issues?
The dissertation included a number of assumptions; the first assumes the relation of demographic variants (gender, age, social status, education, working status
Abstract
This study aimed to survey fungi associated with the product Indomie and Chips being the trades Iargely by a very important segment of society who are the children, beside consumed by adults, but less so, as the survey results to accompany some fungui samples sterile showed proportions presence included various fungi like. Aspergillus flavus, Aspergillus niger, Penicillium Spp., Fusarium graminearum, F.moniliforme, Alternaria alternate and Rhizopus Spp., and other fungi sterile are not diagnosed. The results showed large dominion fungi A. niger by presence sterile samples of both producers, followed by infection in Fusarium Spp., Penicillium Spp., and A. alternata by infection percentage 55, 20 and 17% respectively for the pr