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 novel groups of organic chromophores containing triphenylamine (TPA) (ATP-I to ATP-IV) have been constructed by structural modification of electron donors with substitution biphenyl and bipyridine rings inserting a π-linkage. Density functional theory (DFT) and time-dependent type of it (TD-DFT) have been operated to study results of donating ability of TPA and spacer on absorption, geometrical, photovoltaic, and energetic attributes of these sensitizers. Structural attributes have been revealed that incorporation of TPA, acceptor and π bridge include a perfect coplanar conformation in TPA-III. Based on frequency computations and ground-state optimization, bandgap (Eg) energy, ELUMO, EHOMO have been determined. For enlightening maximu
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Escherichia coli (E. coli) is a frequent gram-negative bacterium that causes nosocomial infections, affecting more than 100 million patients annually worldwide. Bacterial lipopolysaccharide (LPS) from E. coli binds to toll-like receptor 4 (TLR4) and its co-receptor’s cluster of differentiation protein 14 (CD14) and myeloid differentiation factor 2 (MD2), collectively known as the LPS receptor complex. LPCAT2 participates in lipid-raft assembly by phospholipid remodelling. Previous research has proven that LPCAT2 co-localises in lipid rafts with TLR4 and regulates macrophage inflammatory response. However, no published evidence exists of the influence of LPCAT2 on the gene expression of the LPS receptor complex induced by smooth or rough b
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The electrode in the microbial fuel cell has a significant effect on cell performance. The treatment of the electrode is a crucial step to make the electrode surface more habitable for bacteria growth, thus, increases the power production as well as waste treatment. In the current study, two graphite electrodes were treated by a microwave. The first electrode was treated with 100W microwave energy, while the second one was treated with 600W microwave energy. There is a significant enhancement in the surface of the graphite anode after the pretreatment process. The results show an increase in the power density from 10 mW/m2 to 15 mW/m2 with 100w treatment and to 13.47 mW/m2 with 600w treatment. An organic
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In recent years and decades, there is a great need for developing new alternative energy sources or renewable sustainable energy. On the other hand, new technology approaches are growing . towards benefits from the valuable nutrients in wastewater which are unrecoverable by traditional wastewater treatment processes. In the current study, a novel integrated system of microbial fuel cell and anoxic bioreactor (MFC-ANB) was designed and constructed to investigate its potential for slaughterhouses wastewater treatment, nitrogen recovery, and power generation. The system consisted of a double-chamber tubular type MFC with biocathode inoculated with freshly collected activated sludge. The MFC-ANB system was continuously fed with real-fi
... Show MoreBackground: Oral SCC is a complex malignancy where environmental factors, viral infections and genetic alterations most likely interact, and thus give rise to the malignant condition. The HSP70 play a direct role in apoptosis inhibition by aligning the improved integrity of a cell’s proteins with the improved chances of that particular cell’s survival.P21 gene produces p21 protein which is a potent cyclin-dependent kinase inhibitor that plays a significant role in carcinogenesis. The aims of the study were to evaluate and compare the immun-histochemical expression of the HSP70 and cell cycle protein p21in NOM, OED, and OSCC. Correlate both marker expressions with each other. Materials and methods: Forty six formalin-fixed, par
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This work presents a completely new develop an analyzer, named NAG-5SX1-1D-SSP, that is simple, accurate, reproducible, and affordable for the determination of cefotaxime sodium (CFS) in both pure and pharmaceutical drugs. The analyzer was designed according to flow injection analysis, and conducted to turbidimetric measurements. Ammonium cerium nitrate was utilized as a precipitating agent. After optimizing the conditions, the analysis system exhibited a linear range of 0.008-27 mmol. L-1 (n=29), with a limit of detection of 439.3 ng/sample, a limit of quantification of 0.4805 mg/sample, and a correlation coefficient of 0.9988. The repeatability of the responses was assessed by performing six successive injections of CFS at concentra
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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
... Show MoreThis 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
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