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 faujasite type Y zeolite catalyst was prepared from locally available kaolin. For prepared faujasite type NaY zeolite X-ray, FT-IR, BET pore volume and surface area, and silica/ alumina were determined. The Xray and FT-IR show the compatibility of prepared catalyst with the general structure of standard zeolite Y. BET test shows that the surface area and pore volume of prepared catalyst were 360 m2 /g and 0.39 cm3 /g respectively.
The prepared faujasite type NaY zeolite modified by exchanging sodium ion with ammonium ion using ammonium nitrate and then ammonium ion converted to hydrogen ion. The maximum sodium ion exchange with ammonium ion was 53.6%. The catalytic activity of prepared faujasite type NaY, NaNH4Y and NaHY zeolites
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Acid dissociation constants of some Schiff bases derived from 4, 6-dimethyl 2-amino pyrimidine of the type (1) in 50% V/V dioxane-water mixture in 0.003M KCl, at three different temperatures were determined potentiometrically. The thermodynamic energies were calculated and a good linear correlation was obtained between pKa and IR OH. Stretching frequencies.
Were collected three types of medicinal plants from their natural habitat after Astkhalasalziot volatile manner steam distillation and determine the quality and quantity of vehicles chemical for each of the oils obtained using a technique JC discouraged when you merge oily thyme and lemon grass against bacteria either when using oils in three did not have a different effect
Bacteria strain H8, which produces high amount of exopolysaccharide (EPS), was isolated from soil, and identified as strain of Azotobacter chrococcum by its biochemical /physiological characteristics, EPS was extracted, partially purified and used as bioflocculant. The biochemical analysis of the partially purified EPS revealed that it was an alginate. analysis of EPS by Fourier transform infrared spectrometry (FTIR) show that the -OH groups present in bioflocculant are clearly seen at 3433.06 cm-1, the peaks attributed to the -CH3 groups present at 2916.17 cm-1 , and some distinct peaks such as carboxyl group showed strong absorption bands at 1604.66 cm-1, 1411.80 cm-1 and 1303.79 cm-1 indicate the chemical structure of alginate. The effe
... Show MoreThis research aims to identify the factors effecting the investigation of insurance fraud from the perspective of the National Insurance Company employees, by analyzing the study variables and make recommendations on how to enhance the company and employees performance, and to achieve the objectives of the study and test hypotheses a questionnaire was designed after referring to the international standards and the 1969 Iraqi punishment law, and it was answered by 20 employees that were approved for the purposes of this research and analysis.
The study found that lack of employee training was a strong factor, while the supervisory staff and operational procedures was a weak one.
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... Show MoreBackground: Staphylococcus spp. are widely distributed in nature and can cause nosocomial, skin infections, and foodborne illness, and it may lead to severe financial losses in birds by causing systemic infection in numerous organs. Aim: This study was conducted to determine the prevalence of Staphylococcus spp. in humans and birds in Baghdad city. Methods: Seventy-six oral cavity swabs were collected, including 41 from birds and 35 from breeders. All samples were examined by bacteriological methods and identified by using the VITEK technique, the samples were then further studied to test the ability of biofilm formation, and MDR factors and MAR index were tested with the use of seven antibiotics. Results: Among the 76 oral swa
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This study examined the effect of essential oils extracted from peel of Citrus paradisi and Citrus sinensis on two species of fungi: Penicillium oxalicum and Fusarium oxysporum as well as effect of two fungicides: Carbendazim and Thiophanatemethyl against above fungi. Results showed that the essential oil of Citrus paradisi inhibited the radial growth of Penicillium oxalicum and Fusarium oxysporum at concentration 4%. Nevertheless, the essential oil of Citrus sinensis inhibited the radial growth at concentration 5 and 4%, respectively. Furthermore, the two studied fungicides inhibited radial growth of these fungi too. Therefore, there are a positive relationship between the evaluating of concentration and the percentage of inhibiting of rad
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المستودع الرقمي العراقي. مركز المعلومات الرقمية التابع لمكتبة العتبة العباسية المقدسة