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.
A new way to Systems concentrates have been clarified and that allows a concentration high and analysis to automatically wavelengths of the spectrum of this system analyst of the spectrum and the center is built on Holucram Nafez gives less absorbency with efficient diffraction high when the wavelength (900 nm), which will be useful for Khallaya solar
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KE Sharquie, SA Al-Meshhadani, AA Al-Nuaimy, Saudi medical journal, 2007 - Cited by 9
One of these plants utilized in traditional medicine is Lactuca seriolla Linn., which belongs to the Asteraceae family. It goes by a variety of common names in the world, including prickly lettuce, wild lettuce, jagged lettuce, and Kahu and khas. The work aimed to isolate and characterize some bioactive constituent(s) from the aerial part of Lactuca serriola utilizing Combiflash NEXTGEN and high-performance liquid chromatography (HPLC). Lactuca serriola (aerial part) was extracted with 80% ethanol, then fractionated with hexane. Then 250 mg of hexane extract was mixed with 4 g of silica gel and loaded in cartilage, then bounded to the gold column of combi flash using a solvent system comprised of ethyl acetat
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The aim of this study is to utilize the electromembrane extraction (EME) system as a manner for effective removal of zinc from aqueous solutions. A novel and distinctive electrochemical cell design was adopted consisting of two glass chambers, a supported liquid membrane (SLM) housing a polypropylene flat membrane infused with 1-octanol and a carrier. Two electrodes were used, a graphite as anode and a stainless steel as cathode. A comprehensive examination of several influential factors including the choice of carrier, the applied voltage magnitude, the initial pH of the donor solution, and the initial concentration of zinc was performed, all in a concerted effort to ascertain their respective impacts on the efficiency of zinc elim
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A microbial study conducted for a number of flour samples (30 samples) Uses in the bakery ovens in various areas of the city of Baghdad, by used the conventional methods used in laboratories in microbial tests and compared with the modern techniqueby usedof BacTrac Device 3400 equipped from SY-LAB Impedance analysersAustrian company.The results of two ways showed (The conventional way and BacTrac Device test)that the total counts of aerobic bacteria, coliform bacteria, StaphylococcusSpp. bacteria, Bacillus cereus bacteria and yeasts and molds,Most of them were within the permissible borders in the Iraqi standard for grain and its products With free samples from SalmonellaSpp. bacteria, and that the screening by BacTrac device are shorten
... Show MoreLactobacillus Plantarum and Lactobacillus rhamnosus GG were encapsulated using 3% of alginate via extrusion technique. And the probiotics capsules produced were further coated used 1% chitosan to increase the survival of probiotics, and evaluation of The heat resistance of the slow pasteurization and fast pasteurization for Lb,pla and Lb.GG for control and bacteria coated one layer and bacteria coated two layer at 63°C/ 30 minutes and 72°C/ 15 seconds. The results indicate that the Probiotic coated two layer are more resistant to pasteurization temperatures at 63°C/ 30 minutes and 72°C/ 15 seconds than the Probiotic coated one layer. While the results of the control follow a significant reduction for viability of cell toward pasteuri
... Show MoreThe wastewater arising from pulp and paper mills is highly polluted and has to be treated before discharged into rivers. Coagulation-flocculation process using natural polymers has grown rapidly in wastewater treatment. In this work, the performance of alum and Polyaluminum Chloride (PACl) when used alone and when coupled with Fenugreek mucilage on the treatment of pulp and paper mill wastewater were studied. The experiments were carried out in jar tests with alum, PACl and Fenugreek mucilage dosages range of 50-2000 mg/L, rapid mixing at 200 rpm for 2 min, followed by slow mixing at 40 rpm for 15 min and settling time of 30 min. The effectiveness of Fenugreek mucilage was measured by the reduction of turbidity and Chemical Oxygen Demand
... Show MoreAluminum doped zinc selenide ZnSe/n-Si thin films of (250∓20 nm) thickness with (0.01, 0.02 and 0.03), are depositing on the two type of substrate (glass and n-Si) to manufacture (ZnSe/n-Si) solar cell through using thermal vacuum evaporation procedure. physical and optoelectronic properties were examined for the samples. X-Ray and AFM techniques are using to study the structure properties. The energy band gap of as-deposited ZnSe thin films for changed dopant ratio were ranging from (2.6-2.68 eV). The results of Hall effect show that pure and doping films were (p-type), and the concentration carriers and the carriers mobility increases with increase Al-dopant ratio. The (C-V) have shown that the heterojunction were of abrupt type. In add
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