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.
The optimum cultural conditions for garamicidin production by local isolate B.brevis were studied.Best result was obtained when the isolate B.brevis was grown on media composed of 1%glucose as carbon source,1% ammonium chloride as a nitrogen source ,0.5% Dipotassium hydrogen orthophosphate as a phosphate source and after 48 hours of incubation at 30C .Garamicidin has been extracted and purified through acid precipition and then extracted by organic solvent (ether& acetone ).Using HPLC the garamicidin antibiotic showed three types A,B and C garamicidin .
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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Study Purpose. highlighting the origins of the sport's philosophy. Studying the development of the sport of kendo across different historical periods and its adaptation to societal and cultural changes. Identifying the establishment of the most prominent organizations and federations that contributed to its development, and exploring the sport's basic tools and rules and its most prominent championships. Materials and Methods. The historical approach was adopted in studying the historical process of the development of kendo, as it is appropriate to the nature of the problem. The information has been simplified and presented in a sequential manner that presents the development of the sport of kendo, starting from the origins of its p
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Piroxicam is a non-steroidal anti-inflammatory drug (NSAID) used in the treatment of musculo-skeletal and joint disorders. The problem with this drug is its poor solubility in water and hence poor bioavailability after oral administration. In order to improve its solubility and dissolution behavior, hydrophilic additives such as starch, lactose, superdisintegrants including crospovidone (C.P), cross carmellose sodium (CCS), and sodium starch glycolate (SSG) were physically dry mixed with the drug by simple trituration. The improvement in the solubility in 0.1 N HCl was obtained as the amount of starch or lactose increased in the physical mixture, while for superdisintegrants, they further improve the solubility when they are present in s
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Effect of Chlorococcum humicola alcoholic algae extract was studied on the growth of, Pseudomonas aeruginosa, and Klebsiella pneumonia, which were isolated from contaminated water. The extract of Ch. humicola showed a high efficiency in reducing the numbers of the two types of bacteria. . The removal rate of K. pneumonia were 0.0, 48.4 and 57.0, The removal rate of P. aeruginosa were 63.1, 79.8 and 82.9% after24,48, 72 h respectively. The results improved that the K. pneumonia is more sensitive than P. aeruginosa for algae extract concentrations used in study ,and the beast effective time is 24h for the two bacterial species The aim of the study was to eliminate microorganisms using the Alcoholic algae extract. Especially P. aeruginosa and
... Show MoreAmoxicillin 1 was treated with thiosemicarbazide and Phosphoryl chloride to obtain a new derivatives that contains 1,3,4-thiadiazole moiety 2. Schiff bases compounds were synthesized by the reaction of compound 2 with different aldehydes such as benzaldehyde and some substituted Benzaldehyde; p-hydroy, p-Chloro, p-Nitro, p-Dimethylamino, p-Methyl, p-Methoxy, p-Ethoxy to give compounds 3a-h. The obtained compounds have tested towards gram -ve and gram +ve bacteria. The compound shows good to moderate result towards the bacteria.
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