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 research aims to know the traits or characteristics of woman in terms of her external
appearance, motives for her behavior, feelings, mood and ability.
It, moreover, seeks woman’s relationship with others as it is presented by the Iraqi
satirical television show “ State of Melon “.
The researcher adopted for that survey approach using the method of content analysis
to study the research sample represented by “ State of Melon “ show which was
exposed through the screen of a group of channels:
“Hona Baghdad Satellite Channel, then Asia Satellite Channel, Dijla Satellite Channel,
and UTV Satellite Channel.”For this, the researcher used Margaret Gallagher’s Model to analyze the image of
woman in
Out of 150 clinical samples, 50 isolates of Klebsiella pneumoniae were identified according to morphological and biochemical properties. These isolates were collected from different clinical samples, including 15 (30%) urine, 12 (24%) blood, 9 (18%) sputum, 9 (18%) wound, and 5 (10%) burn. The minimum inhibitory concentrations (MICs) assay revealed that 25 (50%) of isolates were resistant to gentamicin (≥16µg/ml), 22 (44%) of isolates were resistant to amikacin (≥64 µg/ml), 21 (42%) of isolates were resistant to ertapenem (≥8 µg/ml), 18 (36%) of isolates were resistant to imipenem (4- ≥16µg/ml), 43 (86%) of isolates were resistant to ceftriaxone (4- ≥64 µg/ml), 42 (84%) of isolates were resistant to ceftazidime (1
... Show MorePolymer metal complexes of poly ethylene glycol acetal and Ag (I), Cu (II), Ni (II), Mn (II), Co (III) and Hg (II) were prepared from the reaction of PEG with aldehyde derived from Erythro-ascorbic acid (pentulosono-ɣ-lactone-2, 3- enedianisoate). All these compounds were characterized by Thin Layer Chromatography (TLC) and FTIR spectra and aldehyde was also characterized by (U.V-Vis), 1HNMR,13CNMR, and mass spectra. It has been established that, the polymer and its metal complexes showed good activities against four pathogenic bacteria (Escherichia coli ,Klebsiellapneumonae, Staphylococcusaureus, Staphylococcus Albus) and two fungal (Aspergillus Niger,Yeast). The polymer metal complexes showed higher activity than the free polymer. The
... Show MoreThe Mannich base ligand was synthesized in an ethanol medium through a condensation reaction of 2-mercaptobenzimidazole and ciprofloxacin at room temperature. Subsequently, several metal complexes of this ligand were prepared. To characterize both the base ligand and the metal complexes, various techniques were employed, including elemental analysis, FT-IR spectroscopy, UV-Vis spectroscopy, molar conductivity measurements, magnetic moment determination, and melting point analysis. The results were shown that the metal complexes formed have the formula [Cr(L)2Cl2] Cl.H2O and [Rh(L)2(H2O)2] Cl3.H2O, where L= mannich base ligand. Based on spectroscopic analytical, coordination with metal ions involves the 'N' donor atom of mannich base
... Show MoreOne of the most effective ways of managing solid waste is microwave-assisted pyrolysis. This research used MW-assisted catalytic co-pyrolysis to extract essential oils from algae powder (AP) and date seed (DS). Graphite and a CaO nanocatalyst were utilized. As for the feedstock, it was a 1:1 mixture of AP and DS. The char content, oil production, and gas generation of co-pyrolysis yields varied between 18.8 wt% and 24.31 wt%, 43.08 wt%, and 55.295 wt%, and 25.905% to 32.60 wt%, respectively. The effect of the CaO nanocatalyst on product yields, feedstock conversion, and heating rates was analyzed. Product yields, average heating rates, and conversion factors were among the metrics studied to determine the synergistic effects and pyr
... Show MoreIn this work, new compounds containing 1,2,4-oxadiazole units were synthesized, characterized, and their crystalline liquid qualities were also studied. The synthesis of compound [I ] starts with the reaction 4-hydroxybenzonitrile with phthaloyl chloride to yield nitrile compound. Then the later compound reacted with hydroxyl amine hydrochloride to obtain compound [II]. Next, 1,2,4-Oxadiazole compounds [III]a-c and [IV] were obtained by the reaction between compound [II] with various acid chlorides(acetyl chloride, chloroacetyl chloride, benzoyl chloride) and phthaloyl chloride in pyridine, respectively. FTIR, 1HNMR and mass spectroscopy were used to analyze the synthesized compounds Spectroscopic investigation verified the synthetic
... Show MoreDate stones were used as precursor for the preparation of activated carbons by chemical
activation with ferric chloride and zinc chloride. The effects of operating conditions represented
by the activation time, activation temperature, and impregnation ratio on the yield and adsorption
capacity towards methylene blue (MB) of prepared activated carbon by ferric chloride activation
(FAC) and zinc chloride activation (ZAC) were studied. For FAC, an optimum conditions of 1.25
h activation time, 700 °C activation temperature, and 1.5 impregnation ratio gave 185.15 mg/g
MB uptake and 47.08 % yield, while for ZAC, 240.77 mg/g MB uptake and 40.46 % yield were
obtained at the optimum conditions of 1.25 h activation time, 500