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.
Achieving goals effectively reflects the success of the institution. However, unless this indicator is coupled with efficiency when achieving goals, the institution will be equal in its achievements, and distinction will remain unachieved. Perhaps the role of the teaching staff in pushing the institution or college towards brilliance focuses on their ability to motivate people on the one hand and their interest in achieving brilliance for the institution. On the other hand, the importance of the research lies in the institution’s reaching a prominent position through the brilliance and creativity of teaching and achieving competition between institutions that make it more brilliance. The study seeks to achieve the goal of the real
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Kinetic experiments were performed to induce of the green methyl dye adsorption from aqueous solution on the bauxite clay. This study includes determination of the adsorption capacity of bauxite clay to methyl green dye adsorption and study the effect of some parameters ( temperature , time ) on the kinetic of the adsorption process of the dye were studied. Quantity of dye adsorbed was increased when the temperature increases from 298 to 318K which indicates that methyl green adsorption processes are endothermic nature . In order to describe the kinetic data and the rate adsorption constants of the pseudo-first-order and second-order kinetics were used . The kinetics data were applied well
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The present paper deal with the issue of the beginning of the culturally
renaissance in emirates of Arab Gulf from 1914-1945 between tow world war
has been attracting the attention of academic about the developments in many
fields in the Arab Gulf at this time.
The paper is divided into five sections. First section, deals with the
geographic importance for the Arab Gulf region. Second section, the economic
situations in the region before and after oil. The third section, talk for social
situations, like population, tribe and tribes in society, and immigration. The
fourth section, deals with the factors of rise the culture and political in the Arab
Gulf before discovery of oil period. The five section, the cultu
In this work the radioactive wastes in the Old Russian
Cemetery Al -Tuwaitha site were classified according to risks for
workers who are involved in the retrieval process. The exposure
assessment results expressed as estimates of radionuclide intakes by
inhalation and ingestion, exposure rates and duration for external
exposure pathways, and committed effective dose equivalents to
individuals from all relevant radionuclides and pathways. Results
showed the presence of natural radionuclides Ra-226, Th-234 and K-
40, as well as the produced radionuclide Cs-137 and Eu-152 in the
cemetery wells. The absorbed doses from the waste were classified to
two categories; exempt waste and low level waste according to
The adsorption study of thymol, was carried out at (25±0.1) °C, using granulated surfactant modified Iraqi Na – montmorillonite clay (initiated modified bentonite); in a down-flow packed column, the modified mineral was characterized by FT-IR spectroscopy. A linear calibration graph for thymol was obtained, which obey Beer's law in the concentration range of 5-50 mg/L at 274 nm against reagent blank. Single-factor-at-a-time approach; showed that the equilibrium time required for complete adsorption was 45 minute with flow rate (4.0drop/ mint). The adsorption of thymol increased with rising pH of the adsorbate solution, increase of solute uptake when the initial adsor
... Show MoreCoagulation - flocculation are basic chemical engineering method in the treatment of metal-bearing industrial wastewater because it removes colloidal particles, some soluble compounds and very fine solid suspensions initially present in the wastewater by destabilization and formation of flocs. This research was conducted to study the feasibility of using natural coagulant such as okra and mallow and chemical coagulant such as alum for removing Cu and increase the removal efficiency and reduce the turbidity of treated water. Fourier transform Infrared (FTIR) was carried out for okra and mallow before and after coagulant to determine their type of functional groups. Carbonyl and hydroxyl functional groups on the surface of
... Show MoreIn the present research the flame retardancy to buildings and industrial foundations which are manufacturing from advanced polymeric composite material was increased by coating it with surface layer included flame retardant material. A(3mm) thick antimony tetroxide was used as a coated layer to retard and prevent the flame spread to the coating surface of polyester resin (SIROPOL 8340-PI) reinforced with hybrid fibers as a woven roving (°45-°0) consist of carbon and kevlar (49) fibers, and exposed it to direct flame generated from gas torch at temperature of (2000ºC), at different exposed distance (10,15,20mm)and study the rang of resistance for this layer and its ability to protec
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