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.
There are numerous ways to define the traits of influential professors, including behavior (such as warmth, civility, and clarity), knowledge (of subject matter and students), and beliefs, among many others. According to research; influential teaching can also defined as the capacity to improve student achievement. This; is only one way to express effectiveness, as stated. However; the preferred definition of high-quality teaching in the United States and many other countries is teacher effects on student achievement. Therefore; this study aimed to determine the characteristics of influential professors in Online Teaching at the English department. This study; believed to be significant for college professors who will show them how
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The finding of novel pollutants in a multitude of surface water bodies worldwide has recently caused environmentalists to face challenges. Therefore, the development of low-cost, efficient technologies is required to provide a safe, pollution-free water environment. The attapulgite surface was used in this study because it has a good absorbent surface for removing bromocresol purple dye from its aqueous solutions. Because attapulgite has a high absorption capacity, it can be used to remove pollutants in both its normal and modified states. Factors affecting adsorption were studied, such as the weight of the adsorbent surface, the initial concentration of the adsorbent, contact time, and the effect of temperature. Clay was ground, cleaned mu
... Show MoreThis 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 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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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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The current research aims to: 1- Identify the emotional sensitivity of children from the parents' point of view. 2- Identifying the differences in the emotional sensitivity of children from the parents ’point of view, according to the parents’ gender variable (father - mother). The basic research sample consisted of (285) male and female students from the sixth grade (primary school), chosen in a randomized stratified method from the districts (Al-Karkh and AlRasafa). While the sample of statistical analysis amounted to (200) father and (200) mothers of male and female students who were randomly selected from the sixth grade of the education directorates (Al-Karkh and Al-Rasafa). The two researchers also used a number of sta
... Show MoreSolanum americanum is a new annual shrubby plant seen recently in fields and gardens of Baghdad city. A new species is described and illustrated, inhabit wet or semi dry places and have consequently a mesophytic habit. A detailed morphological study of the stems, leaves, Inflorescence, flower, male and female reproductive organs and fruits has been done, revealed several interesting taxonomic characteristics, which have not previously been studied in Iraq. Also, anatomical studies reveals constant taxonomical characteristics such as the presence of anthocayanine in outer row of epidermis, distinct chlorenchyma in whole cortex, the wide pith of stems, and presence of distinct mesophyll that differentiated into palisade layer and spongy laye
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Pharmaceuticals are widely distributed in different applications and also released into the environment. Adsorption of Ciprofloxacin HCl (CIPH) on Porcelinaite was studied at ambient conditions. The adsorption isotherms can be well described using the Freundlich and Temkin equations. The pH of the solution influences significantly the adsorption capacity of Porcelinaite, the adsorption of CIPH increased from the initial pH 1.3 and then decreased over the pH rang of 3.8-9. The adsorption is sensitive to the change in ionic Strength, which indicate that electrostatic attraction is a significant mechanism for sorption process. The enthalpy change (∆H) for the adsorption of CIPH onto Porcelinaite signifies an endothermic adsorption. The ∆G
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