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.
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 . &
... Show More
(1)
This study was aimed to investigate the effect of essential oil extracted from the yellow peels of Citrus aurantium on the growth of four species of fungi: Penicillium expansum, Penicillium oxalicum, Fusarium oxysporum and Fusarium proliferatum and effect of one fungicide: Aliette (fosetyl-aluminum) against these fungi. The results showed that the essential oil of C. aurantium inhibited the radial growth of P. oxalicum at concentration 4.5% while P. expansum and F. oxysporum at concentrations 5% and F. proliferatum at concentrations 5.5% additionally the one fungicide tested showed inhibitory effect on radial growth of these fungi. So that there is a negative relationship between the increasing of concentration and radial growth of fungi.
In this work, silver nanoparticles (AgNPs) were biosynthesized from leaves of Ziziphus mauritiana Lam. jujube plant in Iraq and tested against fungal pathogens. Extract of leaves of Z. mauritiana mixed with 10-3 M AgNO3exposed to slight sunlight for 3 days. Characterization of AgNPs was done using UV-visible spectroscopy, SPM (scanning probe microscopy) and atomic force microscopy (AFM). The change of solution color from pale brown to dark brown and the exhibited maximum peak at 445 nm accepted as an indicator to biosynthesized AgNPs. Aqueous extract of Ziziphus mauritiana is considered as biological reduced and stabilized agent for Ag+ to Ag0. AFM showed the formation of irregular shapes of AgNPs. The biosynthesized silver nanoparticles ha
... Show More
(29)
(7)
Aim: Rats are accused in disseminating many zoonotic diseases. This study aimed to isolate and identify bacteria from internal organs of rats captured in Baghdad City, Iraq. Materials and Methods: A total of 120 black rats (R. rattus) were trapped from different areas in Baghdad city. Rats were kept in individual plastic cages for 3 h before euthanizing. Deep pharyngeal swab, intestinal content, urine, and pieces of the liver and spleen, lung, kidney, and brain were obtained aseptically. The specimens were inoculated into peptone water and incubated at 37°C for 24 h for enrichment. A loopful of each specimen was then subcultured onto MacConkey Agar, Blood Agar, and Mannitol Salt Agar. CHROMagar O157 H7 and CHROMagar Listeria were u
... Show More
(4)
Background: The microbial production of substances that have the potency to suppress the growth of other microorganisms is probably one of the prevalent defense strategy developed in nature, microorganisms produce a variable bunch of microbial defense systems, which include antibiotics, metabolic by-products, lytic agents, bacteriocins and others. Objective: The purpose of the present study was to isolate and identify Enterococcus faecium isolates then detecting its ability of carrying the gene responsible for enterocin production in this species. Materials and methods: Out of 50 samples from different sources (food and clinical sources) were collected for the Enterococcus faecium isolation, and the isolated bacteria Enterococ
... Show More
(1)
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 .