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.
The aim of the work is synthesis and characterization of bidentate ligand [3-(3-acetylphenylamino)-5,5-dimethylcyclohex-3-enone][HL], from the reaction of dimedone with 3-amino acetophenone to produce the ligand [HL], the reaction was carried out in dry benzene as a solvent under reflux. The prepared ligand [HL] was characterized by FT-IR, UV-Vis spectroscopy, 'H, 8C-NMR spectra, Mass spectra, (C.H.N) and melting point. The mixed ligand complexes were prepared from ligand [HL] was used as a primary ligand while 8-hydroxy quinoline [HQ] was used as a secondary ligand with metal ion M(IT).Where M(IT) = (Mn ,Co ,Ni ,Cu ,Zn ,Cd and Pd) at reflux ,using ethanol as a solvent, KOH as a
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Recently, there has been an increase in the prevalence of ulcerative colitis (UC), and inflammatory bowel diseases (IBD) worldwide, especially in certain recently industrialized countries like China and In¬dia. Globally, the prevalence of UC, a chronic illness that affects the large intestine, is rising. Fifty adherent invasive Escherichia coli (AIEC) isolates were identified from ulcerative colitis biopsy samples originating from the Gastrointestinal tract (GIT) and Hepatology teaching hospitals/medical city in Baghdad City. The test’s results demonstrated that the AIEC isolates had a high level of resistance to the majority of the an-tibiotics under investigation. Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR) and m
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Abstract: The international community now places significant emphasis on achieving zero carbon emissions, requiring both new researchers and experienced policymakers to prioritise this goal. This article examines the effects of carbon taxes, carbon cap and trade, renewable energy (RE) production and consumption, and economic growth (EG) on carbon emission reduction in the United States, Japan, Canada, and Australia. The study collected secondary data from the World Development Indicators (WDI) secondary source spanning the years 1991 to 2022. The study examines the relationship between variables using the cross-sectionally augmented autoregressive distributed lag (CS-ARDL) approach. The findings indicate that carbon taxes, carbon cap and tr
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In this work, solid random gain media were fabricated from laser dye solutions containing nanoparticles as scattering centers. Two different rhodamine dyes (123 and 6G) were used to host the highly-pure titanium dioxide nanoparticles to form the random gain media. The spectroscopic characteristics (mainly fluorescence) of these media were determined and studied. These random gain media showed laser emission in the visible region of electromagnetic spectrum. Fluorescence characteristics can be controlled to few nanometers by adjusting the characteristics of the host and nanoparticles as well as the preparation conditions of the samples. Emission of narrow linewidth (3nm) and high intensity in the visible region (533-537nm) was obtained.
This study presents a detailed morphology and taxonomic study of Polysiphonia subtilissima collected from Abdul Rehman Goth, Karachi coast, Pakistan. Polysiphonia is a filamentous heterotrichous red algae, characterized by its branching structures and attachment mechanisms. P. subtilissima is notable for its broad salinity tolerance and wide distribution across marine and freshwater ecosystems. This research provides an in-depth examination of the internal and external structures of P. subtilissima, contributing to its systematic study and documenting its first recorded occurrence in Pakistani coastal areas, bordering the northern Arabian Sea. The findings enhance the understanding of the species taxonomy and its ecological role in
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Fresh vegetables are an important part of a healthy diet. The consumption of raw vegetables without cooking or good washing can be a major rout of transmission to the parasitic infection. The goal of this study was to determine the intestinal parasitic contamination of fresh vegetables from vegetables sales markets in Baghdad province during the different above months of the year. A total of 303 samples of different vegetables were randomly selected from three wholesale markets distributed through different regions in Baghdad (East, West and South) and then were examined by a floatation method. The present study showed that the collected vegetables were contaminated with 12 species of intestinal parasites, and the total percentage of contam
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The purpose of my thesis is to synthesis two new bidentate ligands which were used to prepare series of metal complexes by reacting the ligands with (M+2 = Mn, Co, Ni, Cu, Cd and Hg) Succinyl chloride was used as starting material to synthesis two bidentate ligands (L1) and (L2) by reaction it with 4-chloroaniline (L1) and (4-aminoacetophenone) (L2) in dichloromethane as a solvent, that are: (L1) = N1,N4-bis (4-chloro phenyl ) succinamide (L2) =N1,N4-bis(4-acetylphenyl)succinamide The new ligands were characterize by using spectroscopic study (Fourier-transform infrared spectroscopy (FT-IR), electronic spectra ( UV-Vis) ,nuclear magnetic resonance(1H,13C-NMR), Mass spectra
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