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 .
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 MoreThe Diyala River is considered the third most important river in Iraq. However, in the recent period, Diyala Governorate has been subject to several floods. This study aims to simulate an efficient labyrinth weir at the flood escape entrance branching from the Diyala River to reach the best entrance through which the flood waves can pass safely. The discharge coefficient was calculated laboratory for five types of trapezoidal side labyrinth weirs with different sidewall angles. Results showed that the coefficient discharge for the trapezoidal labyrinth side weir with an angle of the sidewall is 75ᵒ and has a discharge coefficient greater than the rest of the labyrinth side weirs. The second part of this study is valida
... 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.
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.
The current study was conducted on goats in various parts of Wasit Province, Iraq, from November 2021 to April 2022. The study aims to find and identify intestinal parasites (IPs) in goats in Wasit province. The goat's fresh fecal specimens (n=180) include cysts, eggs, oocysts, trophozoites and larval stages. One hundred eighty sheep feces samples were collected, and more than one parasite was isolated from one sample (mixed infection). According to the data acquired, the overall prevalence of intestinal parasites in goats was 52.77 (95 samples). In the current investigation, eleven distinct (IPs) species with infection rates were identified, including Toxocara vitulorum (Goeze, 1782) (16.66 %), Cryptosporidium sp.( Tyzzer, 1907) (1
... Show MoreAim: 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
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