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.
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The presence of hydrocarbons in the soil is considered one of the main problems of pollution. In our current study, eight samples isolated from soil saturated with hydrocarbons were taken from different areas of Baghdad, Iraq. In this study, 5 isolates belonging to Pseudomonas aeruginosa by 99%, 4 isolates to Klebsiella pneumoniae by 98%, and 3 isolates to Enterobacter hormaechei by 97% were diagnosed in different ways. A molecular examination was also conducted by 16sRNA. We recorded P. aeruginosa, K. Pneumoniae and E. hormaechei as new local isolates in NCBI. In addition, a comparison was made between our isolates and the global isolates to determine the degree of convergence in the evolutionary line. The genes alkB and nahAc7 were diagno
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Oxidation of sulfur compounds in fuel followed by an adsorption process were studied using two modes of operation, batch mode and continuous mode (fixed bed). In batch experiment oxidation process of kerosene with sulfur content 2360 ppm was achieved to study the effect of amount of hydrogen peroxide(2.5, 4, 6 and 10) ml at different temperature(40, 60 and 70)°C. Also the effect of amount acetic acid was studied at the optimal conditions of the oxidation step(4ml H2O2 and 60 °C).Besides, the role of acetic acid different temperatures(40, 60, 70) °C and 4ml H2O2, effect of reaction time(5, 30, 60, 120, 300) minutes at temperatures(40,60) °C, 4ml H2O2 and 1 mlHAC)&
... Show MoreProbiotics are live microbes that give many health benefits to human beings and animals, the most studied and commonly used probiotics are Gram-positive bacteria; lactobacilli and bifidobacteria. At nowadays, Lactobacillus spp. constitute more than two-thirds of the total numbers of probiotic species. The present study aimed to characterize Lactobacillus that locally isolated from human mouth and feces as probiotics. A total of three Lactobacillus isolates; Lactobacillus fermentum Lb2, Lactobacillus rhamnosus Lb9, and Lactobacillus paracasei Lb10 were investigated in respect to acid and bile salts tolerance, antibiotics susceptibility, and cell surface hydrophobicity in vitro using bacterial adhesion to hydrocarbons method. In compa
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Room temperature ionic liquids show potential as an alternative to conventional organic membrane solvents mainly due to their properties of low vapour pressure, low volatility and they are often stable. In the present work, the technical feasibilities of room temperature ionic liquids as bulk liquid membranes for phenol removal were investigated experimentally. In this research several hydrophobic ionic liquids were synthesized at laboratory. These ionic liquids include (1-butyl-3-methylimidazolium bis (trifluoromethylsulfonyl) imide[Bmim][NTf2], 1-Hexyl-3-methylimidazolium bis (trifluoromethylsulfonyl) imide[Hmim][NTf2], 1-octyl-3-methylimidazolium bis (trifluoromethylsulfonyl)imide[Omim][NTf2],1‐butyl
... Show MoreThe adult worms of the Microphallidae family are mainly found as intestinal parasites of birds and mammals, while metacercariae is most commonly found in decapodal crustaceans. The Microphallidaeare family is spread throughout the world. It includes approximately 47 genera. Mature worms usually enter the digestive system of vertebrates, especially birds and mammals. Microphallidae contain eight subfamilies: Androcotylinae - Basantisiinae - Endocotylinae - Gynaecotylinae - Levinseniellinae - MaritrematinaeMicrophallinae - Sphairiotrematinae. Therefore, due to the lack of studies on the Microphallidae family in Iraq, we began to develop a database on this important family.
BACKGROUND: Color Vision Deficiency (CVD) is mostly an inherited trait and is not an uncommon problem. Prevalence of CVD differs among different ethnic and geographic properties of the population that affect their genetic constitution. Ishihara plates remain an internationally accepted tool for screening red-green CVD. OBJECTIVE: To determine the prevalence of red-green CVD among adult males from Baghdad province. PATIENTS AND METHODS: One thousand and five (1005) adult males were enrolled in this study, using a systematic sampling technique, and were screened for CVD utilizing 24-plate Ishihara plates and re-tested by EnChroma 39-Color plates. All males were residing in Baghdad and the center of Iraq. RESULTS: Among all tested males, 948 r
... Show MoreAs a marker of systemic inflammation, raised (C-reactive protein (CRP)) concentrations which are still within the normal range have been associated with an increased inflammation of chronic renal diseases (CRD). The current study aimed to establish potential determinats of raised CRP concentrations in patients who treated in Heamodialysis room,then study the relationship between CRP& some biochemical parameters related CRD We used a CRP latex reagents Kit which is based on an immunological reaction between CRP antisera bounded to the biologically inert latex particles or with CRP in the test specimens of 19 patients with (CRD) mean age 48 years ,range = 30?65 & in 21 healthy subjects as control group their age range = 30 ?45 years. The
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This work was conducted to study the extraction of eucalyptus oil from natural plants (Eucalyptus camadulensis leaves) by organic solvents. the effects of the main operating parameters were studied; type of solvent (n-hexane and ethanol), time to reach equilibrium, the temperature (45°C to 65°C) for n-hexane and (45°C to 75°C) for ethanol, solvent to solid ratio (5:1 to 8:1 (v/w)), agitation speed (0 to 900 rpm) and the particle size (0.5 to 2.5 cm) of fresh leaves to find the best processing conditions for the achieving maximum oil yield. The concentration of eucalyptus oil in solvent was measured by using UV-spectrophotometer. The results (for n-hexane) showed that the agitation speed of 900 rpm, temperature 65°C with solvent to soli
... Show MoreDue to the broad range uses of chromium for industrial purposes, besides its carcinogenic effect, an efficient, cost effective removal method should be obtained. In this study, cow bones as a cheap raw material were utilized to produce active carbon (CBAC) by physiochemical activation, which was characterized using: SEM to investigate surface morphology and BET to estimate the specific surface area. The best surface area of CBAC was 595.9 m2/gm which was prepared at 600 ᵒC activation temperature and impregnation ratio of 1:1.5. CBAC was used in aqueous chromium ions adsorption. The investigated factors and their ranges are: initial concentration (10-50 mg/L), adsorption time (30-300 min), temperature (20-50
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