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.
Schiff base derived from PVA and Erythroascorbic acid derivative (pentulosono-ɣ-lactone-2, 3-enedianisoate) was synthesized and characterized by Thin Layer Chromatography (TLC) and FTIR spectra, aldehyde was also characterized by (U.V-Vis), 1HNMR, 13CNMR and mass spectra. The inhibitory effect of prepared polymer on the activity of human serum Cholinesrerase has been studied in vitro. The polymer showed a remarkable activity at low concentration (4.5*10-3 – 4.5*10-8 M).
Relations of the prophets Muhammad (May peace be upon him) with by
gantlines from the revelation to the trance of al-hdybya 6 A.H/610 A.C.
This research is concerned with Islamic historical events, particularly dunning
the time of the prophet Muhammad. It also explores relations of the apostle of god
with the rums and their allies the Arabs of Syria.
During the first years of revelation Muslim had achieved great victories
against the unbelievers. This accomplishm-ent had draw attention of some foreign
forces such as the Arabs of Syria, who became snapicions of this new and active
power. Their hostile attend was demonstrated by striking an economical siege against
Muslims. They attacked the nabtins merchants who
The present research included synthesis of silver nanoparticle from(1*10-3,1*10-4 and1*10-5) M aqueous AgNO3 solution through the extract of M.parviflora reducing agent. In the process of synthesizing silver nanoparticles we detected a rapid reduction of silver ions leading to the formation of stable crystalline silver nanoparticles in the solution.
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Schiff base ligand (H2CANPT) was prepared by two steps: first, by the condensation of curcumin with 4-amino antipyrin produces4,4'-(((1E,3Z,5Z,6E)-1,7-bis(4-hydroxy-3- methoxyphenyl)hepta-1,6-diene-3,5-diylidene)bis(azanylylidene))bis(1,5-dimethyl-2-phenyl- 1,2-dihydro-3H-pyrazol-3-one) (CANP). Second, by the condensation of (CANP) with L-tyrosine produces2,2'-(((3Z,3'Z)-(((1E,3Z,5Z,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta 1,6-diene-3,5-diylidene)bis(azanylylidene))bis(1,5-dimethyl-2-phenyl-1,2-dihydro-3-H-pyrazole- 4-yl-3-ylidene))bis(azanylylidene))bis(3-(4-hydroxyphenyl)propanoic acid) (H2CANPT). The resulted Schiff comported as hexadentate coordinated with (N4O2) atoms, then it was treated with some transition and non-transaction met
... Show MoreThis work deals with the description of histopathological effects of the nematode Hartertia
gallinarurn Theiler. 1919 on the digestive system of the seesee partridge collected from Qa’ra
area in the western desert district of Iraq. along with some notes on intensity fluctuation of the
parasite according to the seasons. It is found that the major effects of the nematode are
necrosis and fibrosis of gizzard: granulomatous reaction. necrosis and mononuclear
infiltration of proventriculus: damage of mucosal lining of intestine and lymphocytic
infiltration of liver.
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Motives for public exposure to specialized sports satellite channels and the gratifications achieved about it - Research presented by (Dr. Dr. Laila Ali Jumaa), Imam Al-Kadhim College (peace be upon him) - Department of Information-2021.
The research aims to know the extent of public exposure to specialized sports satellite channels, and what gratifications are achieved from them, and to reach scientific results that give an accurate description of exposure, motives and gratifications verified by that exposure, and the research objectives are summarized in the following:
- Revealing the habits and patterns of public exposure to specialized sports satelli
This piece of research work aims to study one of the most difficult reaction and determination due to continuous and rapid variation of reaction products and the reactants. As molybdenum (VI) aid in the decomposition of hydrogen peroxide in alkaline medium of ammomia, thus means a continuous liberation of oxygen which cuases and in a continuous manner a distraction in the measurement process. On this basis pyrogallol was used to absorbe all liberated oxygen and the result is an a clean undisturbed signals. Molybdenum (VI) was determined in the range of 4-100 ?g.ml-1 with percentage linearity of 99.8% or (4-300 ?g.ml-1 with 94.4%) while L.O.D. was 3.5 ?g.ml-1. Interferring ions (cations and anions) were studied and their main effect was red
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