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.
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New Schiff bases derived from D-galactose were synthesized by condensation of aldehyde (1,2:3,4-Di-O-isopropylidene-6-carboxaldehyde-α-D-galactopyranose) with different aromatic amines such as (4-bromo, 3-hydroxy, 4-iodo, 4-methoxy) aniline in dry benzene using glacial acetic acid as a catalyst. These compounds were converted to oxazepine derivatives by addition reaction with maleic anhydride in dry benzene as a solvent. The structures of the synthesized compounds have been characterized by elemental analysis, FTIR spectra, some of them by using 1HNMR spectra and measurement of its physical properties.
The presence of dyes in wastewater has become a major issue all over the world. The discharge of dyes in the environment is concerned for both toxicological and esthetical reasons. In this study, the removal of dyes from aqueous solution by electrocoagulation using aluminum electrodes as cathode and anode were investigated with the electrocoagulation cell of 1litter. The study included: the impact of various operating parameters on the dyes removal efficiency like pH, NaCl concentration, distance between electrodes, voltage, initial dyes concentration and type of electrodes. The dye (congo red) concentrations were (50, 100, 150, and 200 ppm), stirring speed was 120 rpm at room temperature. pH used was maintained constant
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The cuneiform texts considered one of the most important resources which play an important role help us to know how was the economic , politic and religious manner and every thing have relation of the history of Mesopotamia which contain many of information that enrich our knowldgement every side of Mesopotamia civilization since using writing as away to register the science and human knowldgement . The city of Sippar (Abu –Habba ) was one of most important city of ancient Mesopotamia which mentioned as one of first five Mesopotamian cities found before floating and it exposed through excavating rich one of them tablet which some of it dated to old Babylon period . The king Ammi-Saduqa has a big share of tablet we study number of thi
... Show MoreThe study's objective is to produce Nano Graphene Oxide (GO) before using it for batch adsorption to remove heavy metals (Cadmium Cd+2, Nickel Ni+2, and Vanadium V+5) ions from industrial wastewater. The temperature effect (20-50) °C and initial concentration effect (100-800) mg L-1 on the adsorption process were studied. A simulation aqueous solution of the ions was used to identify the adsorption isotherms, and after the experimental data was collected, the sorption process was studied kinetically and thermodynamically. The Langmuir, Freundlich, and Temkin isotherm models were used to fit the data. The results showed that Cd, Ni, and V ions on the GO adsorbing surface matched the Langmuir model with correlation coefficients (R2)
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Through his female characters, Walt Disney was able to reflect the
development of the general outlook upon women in the Western world over a period
of sixty years.
This paper sheds light in how the cartoon characters changed in their reaction
to problems and means of solving them from the 30s through to the 90s of the
previous century. The main characters will be seen according to the films’ dates of
production and release.
This research has been prepared to isolate and diagnose one of the most important vegetable oils from the plant medical clove is the famous with Alaeugenol oil and used in many pharmaceuticals were the isolation process using a technique ultrasonic extraction and distillation technology simple
BACKGROUND: Color Vision Deficiency (CVD) is mostly an inherited trait and is not an uncommon problem. Prevalence of CVD differs among different ethnic and geographic properties of the population that affect their genetic constitution. Ishihara plates remain an internationally accepted tool for screening red-green CVD. OBJECTIVE: To determine the prevalence of red-green CVD among adult males from Baghdad province. PATIENTS AND METHODS: One thousand and five (1005) adult males were enrolled in this study, using a systematic sampling technique, and were screened for CVD utilizing 24-plate Ishihara plates and re-tested by EnChroma 39-Color plates. All males were residing in Baghdad and the center of Iraq. RESULTS: Among all tested males, 948 r
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