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
Objective: Zerumbone (ZER) is a well-known natural compound that has been reported to have anti-cancer effect. Thus, this study investigated the ZER potential to inhibit Thymidine Phosphorylase (TP) and the ability to trigger Reactive oxygen species (ROS)-mediated cytotoxicity in non-small cell lung cancer, NCI-H460, cell line. Material and Method: The antiangiogenic activity for ZER was evaluated by using the thymidine phosphorylase inhibitory test. Reactive oxygen species (ROS) production was determined via DCFDA dye by using flow cytometry. Result and Discussion: ZER was found to be potent TP inhibitory with the IC50 value of 50.3± 0.31 μg/ml or 230±1.42 µM. NCI-H460 cells upon treatment with ZER produced sign
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Human interferon as is the case in all kinds of interferon has complex effects but all share their impact on preventing the proliferation of viruses and preventing or reducing human Alantervjørn conversion occurs if the cell is in preventing the growth of the virus when interferon Balnmstqubl connects
This research aims to study the effect of heat on the efficiency of solar cells of neutrons ranging from card to these cells in the case of dark and light before and after irradiation using the neutron source as well as electrical properties have been studied
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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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Lactobacillus 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 MoreA 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
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