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.
A theoretical and protection study was conducted of the corrosion behavior of carbon steel surface with different concentrations of the derivative (Quinolin-2-one), namly (1-Amino-4,7-dimethyl-6-nitro-1H-quinolin-2-one (ADNQ2O)). Theoretically, Density Functional Theory (DFT) of B3LYP/ 6-311++G (2d, 2p) level was used to calculate the optimized geometry, physical properties and chemical inhibition parameters, with the local reactivity to predict both the reactive centers and to locate the possible sites of nucleophilic and electrophilic attacks, in vacuum, and in two solvents (DMSO and H2O), all at the equilibrium geometry. Experimentally, the inhibition efficiencies (%IE) in the saline solution (of 3.5%) NaCl were studied using potentiomet
... Show More
(13)
(4)
The current research aims to investigate the counseling services provided to gifted students in the Al-Baha region from their point of view, and to identify the differences in these views according to the students ’gender, educational stages, and the cities in which they study. The descriptive survey approach was followed in its relevance to the research objectives. Likewise, a random sample of gifted elementary, intermediate, and secondary school students was selected by the method of stratified sampling, as the sample reached (175) male and female students. The instrument of the research consisted of the questionnaire of counseling services presented to gifted students from their point of view prepared by researchers. The results sho
... Show MoreThis note reported the first record of Spotted Flycatcher Muscicapa striata (Pallas, 1764) (Passeriformes, Muscicapidae) from the state of Odisha, India. This species was recorded from the north and western part of the country as well as from the Western Ghats, but this note reports the first record from the Eastern Ghats of India.
Green synthesis of silver nanoparticles (AgNPs) using different plant parts has shown a great potential in medicinal and industrial applications. In this study, AgNPs were in vitro green synthesized using A. graecorum, and its antifungal and antitumoractivities were investigated. Scanning electron microscopy (SEM) image result indicated spherical shape of AgNPs with a size range of 22-36 nm indicated by using Image J program. The functional groups indicated by Fourier-transform infrared spectroscopy (FTIR) represented the groups involved in the reduction of silver ion into nanoparticles. Alhagi graecorum AgNPs inhibited MCF-7 breast cancer cell line growth in increased concentration depend manner, significant differences shown at
... Show More
(137)
(130)
(12)
(10)
In this study lattice parameters, band structure, and optical characteristics of pure and V-doped ZnO are examined by employing (USP) and (GGA) with the assistance of First-principles calculation (FPC) derived from (DFT). The measurements are performed in the supercell geometry that were optimized. GGA+U, the geometrical structures of all models, are utilized to compute the amount of energy after optimizing all parameters in the models. The volume of the doped system grows as the content of the dopant V is increased. Pure and V-doped ZnO are investigated for band structure and energy bandgaps using the Monkhorst–Pack scheme's k-point sampling techniques in the Brillouin zone (G-A-H-K-G-M-L-H). In the presence of high V content, the ban
... Show More
(6)
Biodegradation is utilizing microorganisms to degrade materials into products that are safe for the
environment, such as carbon dioxide, water, and biomass. The current study aims to isolate and characterize
bacteria with polyethylene terephthalate (PET) degradation ability isolated from Shatt al-Arab water and
sewage from Basra, the bacteria were identified as Klebsiella pneumonia. According to the findings, the
isolates showed a highly significant difference in degradation of PET (24% during 7 days) and the percent of
degradation increased to 46% at 4 weeks compared to the control. The study also involved determining the
optimum temperature of K. pneumonia growth, which was 37°C, while the preferred
(5)
Stable isotopes 18O/16O and 13C/12C in the carbonate rocks of the Mishrif Formation are examined here to define the depositional characters in the basin includes paleo temperatures and paleo depth. The Mishrif formation (Cenomanian – Early Turonian) has extensive distribution in Iraq and Middle East. Mishrif Formation composed of organic detrital limestone. Four boreholes in four oilfields, Noor – well (11), Amarah – well (14), Buzurgan – well (24), Halfaya – well (8), in south east of Iraq have been studied. The studied samples have negative δ18O isotope values studied well, with Average (-4.11‰), (-4.47‰), (-4.48‰), (-4.18‰) in the studied wells res
... Show More
(4)
(3)
A batch adsorption system was applied to study the adsorption of methylene blue from aqueous solution by Iraqi bentonite and treated bentonite with different amount of zinc oxide (ZnO). The adsorption capacities of methylene blue onto bentonite were evaluated. The equilibrium between liquid and solid phase was described by Langmuir model better than the Freundlich model. Langmuir and Freundlich constants have been determined. The separation factor or equilibrium parameter, RL which is used to predict if an adsorption system is favourable or unfavourable was calculated for all cases.