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 study aims at identifying the morphological and psycho-socio-economic qualities wished to be in a life partner among a sample of Palestinian youth. The total sample was (231) and consisted of (83) male and (148) female students. Each student presented a detailed report on the qualities he/she wished to be in life partner. The study used the descriptive approach and content analysis method. The validity and stability of the analysis were calculated. The results showed eight qualities in both groups: physical, psychological, emotional, social, intellectual, familial, economic and academic. Female students were found to have more variations than male students in terms of the qualities preferable in the life partner. Male students
... Show MoreNigella sativa has various pharmacological properties and has been used throughout history for a variety of reasons. However, there is limited data about the effects of N. sativa (NS) on human cancer cells. This study aimed at observing the roles of methanolic extract of N. sativa on apoptosis and autophagy pathway in the Human PC3 (prostate cancer) cell line. The cell viability was checked by MTT assay. Clonogenic assay was performed to demonstrate clonogenicity and Western blot was used to check caspase-3, TIGAR, p53, and LC3 protein expression. The results demonstrated that PC3 cell proliferation was inhibited, caspase-3 and p53 protein expression was induced, and LC3 protein expression was modulated. The clonogenic assay showed that PC3
... Show MoreIn recent years, it has been evident that searching for alternative methods with low-price and eco-friendly features that produce high-quality adsorbents is in high demand. In the present work, Rice husk from Iraqi rice named (Amber) had been used as the primary source to produce rice husk ash (RHA) for the removal of the antibiotic metronidazole (Flagyl) from water. After optimum drying of rice husk, rice husk ash (RHA) was obtained at 600 °C using an electric oven. RHA has been investigated for properties using X-ray diffraction (XRD), porosity, and surface area (SA). The experimental work adsorption data were optimized to evaluate Langmuir and Freundlich constants. The thermodynamic parameters likely a change in Gipp's energy (ΔG),
... Show MoreActivated carbon was Produced from coconut shell and was used for removing sulfate from industrial waste water in batch Processes. The influence of various parameter were studied such as pH (4.5 – 9.) , agitation time (0 – 120)min and adsorbent dose (2 – 10) gm.
The Langmuir and frandlich adsorption capacity models were been investigated where showed there are fitting with langmmuir model with squre regression value ( 0.76). The percent of removal of sulfate (22% - 38%) at (PH=7) in the isotherm experiment increased with adsorbent mass increasing. The maximum removal value of sulfate at different pH experiments is (43%) at pH=7.
A process of bacterial cellulose gold nanocomposite has been investigated based on experimental work and cited literature. A literature review on the production process is carried out in this study. Bacterial cellulose is a high crystalline fabric material generally used in biomedical applications. A Nanocomposite was made by synthesis from gold and bacterial cellulose. The experimental work includes growing, and isolating bacterial cellulose, preparation of gold Nanoparticles and preparation of Nano composite. Nanoparticle’s formation and adsorption on the cellulose tissue have been observed visually, where a colour change was observed. The predicted particle size for the gold nano
In the present study, advanced oxidation process / heterogeneous photocatalytic process (UV/TiO2/Fenton) system was investigated to the treatment of oily wastewater. The present study was conducted to evaluate the effect of hydrogen peroxide concentration H2O2, initial amount of the iron catalyst Fe+2, pH, temperature, amount of TiO2 and the concentration of oil in the wastewater. The removal efficiency for the system UV/TiO2/Fenton at optimal conditions and dosage (H2O2 = 400mg/L, Fe+2 = 40mg/L, pH=5, temperature =30oC, TiO2=75mg/L) for 1000mg/L load was found to be 77%.
Aluminum foil cover around the re
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