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.
Abstract
This study aims to evaluated the user satisfaction of retrieval services
concerning to universities thesis and dissertations in university dissertation
unit of Baghdad Library for achieving the following objectives:
1- Evaluating the performance of this unit (thesis unit of Baghdad University
Library) regarding to users opinion.
2- Recognizing the reasons in this unit behind the case of non satisfaction of
its users and trying to find the suitable solutions.
To achieve those two objectives, the questionnaire tool was performed
and determined the user's satisfaction level by using a sample survey. 1118
graduated students were subjected to this experiment. The following main
results were appeared:<
This research involved synthesis of new β-Lactam derivative from Azo compound[4-amino-N-(pyrimidine-2-yl)-3-(pyrimidine-2-yldiazenyl) benzene sulfonamide] (S1) record previously by many steps. Starting conversion the free amino group in an azo comp. to chloro acetamide derivative(S2), then reacted it with urea to give the oxazole ring derivative (S3) that which containing free amino group. The condensation reaction between the amino group and P-bromobenzaldehyde to produce Shiff base (B14). Finally staudinger's cyclo addition reaction go run between the Shiff base derivative (B14) and chloro acetyl chloride in the presence of tri ethyl amine (Et3N) as Base catalyst and dioxane a
... Show MoreThe surplus glycerol produced from biodiesel production process as a by-product with high quantity can be considered as a good source to prepare glycerol carbonate (GC) whereas with each 1000 kg from biodiesel obtains 100 kg from glycerol. Glycerol converted to glycerol carbonate over bio-char as a catalyst prepared by slow pyrolysis process under various temperatures from 400 ᴼC to 800 ᴼC. The char prepared at 700 ᴼC considered as a best one between the others which was manufactured to activate the transesterification reaction. GC have large scale of uses such as liquid membrane in gas separation, surfactants ,detergents , blowing agent , in plastics industry, in Pharmaceutical industry and electrolytes in lithium batteries.
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The objective of present study was to investigate the effect of using mixture volaticle oil of rosmarinus and nigella sativa to improve some of the meat quality characteristics, physical and limited storage time of minced cold poultry meat. Duplex volaticle oil was added at 0.025, 0.050 and 0.075 g/kg to minced poultry meat, these treatments were stored individually for 0 , 4 and 7 days at 4-7C0. After making several chemical, physical and oxidation indicators, the following results were obtained:
The process of adding volaticle oil to minced poultry meat led to significant increase (P<0.01)in moisture, prot
... Show MoreActivity test of the inhibitors purified from barley and broad beans crop proved the inhibition activity against 6 types of rots Pencillium ssp and Aspergellusflavus and Aspergillus niger and Fusarium solani and Fusarium semitectum and Mucor with three concentrations 0.1 and 0.2 and 0.3 mg/ml, where the inhibitor purified from the second peak of broad beans proved that it had a higher inhibition activity against the growth of test rots which were 53.75 and 62.5 and 78.5 and 76.25 and 84 and 18.8% respectively, at 0.3 mg/ ml followed by the first peak of the inhibitor purified from broad beans the inhibition activity were 43.75 and 50 and 62.96 and 75 and 80 and 12.5 then the inhibitor purified from barley in which the inhibition activity
... Show MoreEnvironment suffered in recent years a large corrupting by human; and because of his ignorance of the dimensions of Caliphate in the ground and ignore what it means gearing. The gearing is that the son of Adam, which will benefit the board of Allah Almighty to him in the land of the causes of life, without exaggeration or negligence and without prejudice to the cosmic Balnoames enacted by the Almighty Creator, has urged verses of the Quran Muslim to preserve and protect the environment which is a religious duty, as it showed a great verses he is the author and the splendor and beauty of workmanship and manufacturer greatness of the Almighty, who created all things beautiful.
Koran played a major role in the consolidation of environmen
Information on the scorpions' fauna of Iraq is limited especially in Thi Qar Province. The scorpion specimens of the present study were collected from the desert area which is located between the provinces of Thi Qar and Al-Muthana (Al-Kata'a region). The Scorpio kruglovi (Birula, 1910) redescribed in this study was found in this area. The diagnostic characters are given and the important features are figured.
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The parasite tapeworm (Raillietina echinobothrida) belonges to the class Cestoda, it is responsible for nodular tapeworm disease in poultry .The aim of this study was to determine tapeworm parasites infections in Columba livia from two markets in the province of Baghdad for the period from May to December 2014. From a total of thirty five sample of Columba livia were randomly selected and then examined the elementary canal of these samples. The present study showed that the collected rock pigeon were found six infected with the cestode Raillietina echinobothrida with infection rate (17.14%). The statistical analysis for the characters of the cestode showed significant differences in all recipes, but there were no significant difference
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This research included recent skills of transformed ieadership ,its five aspect and they are(transformed,speech of boos,chorus co-0perative work,behavior adjustment of values,common view point,understanding,and apologizing)Thescale is being made for these five aspects and it done on sample of Baghdad university staff and the sample consist of(424)teacher of both genders.The study found out existence of transformedskills of leadership for girls . &
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