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.
Abstract
The purpose of the present paper is to light on the relationship between jobs design, analysis and its reflections on reinforcing workers' vocational adjustment. The present paper aims to accomplish cognitive and applied goals, top of which, test of functional analysis ability to have effect upon workers' vocational adjustment via job design directly and indirectly owning to the virtual factor practiced by these practices on the sought organization. The problem of the present paper comes with many, the most important is the of how to bolster and back up worker's technical adjustment through good and accurate design for the job.
Based on this problem and goals as to expla
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Objective: The goal of this research was to evaluate where selenium nanoparticles impact the activity of antibodies in immunized lambs with foot and mouth vaccines by modulating the immune system. Materials and Methods: Two groups of lambs of 3–4 months of age were injected with 1 ml of ARRIAH-VAC vaccine intramuscularly in the neck, five Lambs were given selenium nanoparticles (size 100 nm) oral administration of selenium nano dose of 0.1 mg/kg of body mass once every day for sixty days considered as group one (G1) while the other five used as control Group 2 (G2). Results: This resulted in the establishment of an immune response, as evidenced by a rise in antibody titer in the blood using the ELISA test for three serotypes A,
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The main object of the current work was to determine the antifungal efficiency of secondary metabolites product called synephrine that extracted from Citrus sinesis peels and the ability of synephrine to biosynthesis gold nanoparticles from HAucl4 which consider environmentally favourable method, then determine their activity against pathogenic human dermatophyte. The identification of synephrine done by Thin layer chromatography (TLC), High Performance Liquid Chromatography (HPLC) and The Fourier Transform Infrared (FTIR). The characterization of gold nanoparticles by using Ultra Violet-Visible Spectroscopy (UV-Vis), Field – Emission Scanning Electron Microscopy (FESEM) and Fourier Transform Infrared (FTIR), confirmed the biosynt
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Amaranthus viridis L. belongs to the Amaranthaceae family. It is a rich source of numerous phytochemicals and amino acids. The objective of this work was to optimize Ultrasound-Assisted Extraction (UAE) based on the extraction yield and Thin-Layer Chromatography (TLC) profile under different conditions, to compare the optimized UAE to the Soxhlet extraction method and evaluate the cytotoxic effects of the ethyl acetate fraction of the 80 % ethanolic extract on the SKGT-4 (human esophageal adenocarcinoma), AGS (human gastric adenocarcinoma) and A431 (human epidermoid carcinoma). A one-factor at a time experiment was carefully designed to assess the influence of the following factors on the extraction: time, frequency, solid-to-solven
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Abstract The study aimed at reviewing translation theories proposed to address problems in translation studies. To the end, translation theories and their applications were reviewed in different studies with a focus on issues such as critical discourse analysis, cultural specific items and collocation translation.
We have investigated twenty five patients with type-2 diabetes mellitus aged (35-60) years and fifteen healthy persons as control group to detect Anti-Helicobacter pylori IgG antibody. All studied groups were carried out to measure fasting blood sugar, anti- Glutamic acid decarboxylase (GAD), anti-? islets cells antibody by IFAT, Anti-H. pylori IgG antibody by ELISA technique. There was significant elevation in the concentration of fasting blood sugar than in control group (P < 0.05), the patients had negative results for anti-GAD antibody and anti- ? islets cells antibody, there were significant differences (P < 0.05) of anti-H. pylori IgG antibody in 28 % of patients had type-2 diabetes than control group. This lead to suggestion that typ
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In this work, a novel design for the NiO/TiO2 heterojunction solar cells is presented. Highly-pure nanopowders prepared by dc reactive magnetron sputtering technique were used to form the heterojunctions. The electrical characteristics of the proposed design were compared to those of a conventional thin film heterojunction design prepared by the same technique. A higher efficiency of 300% was achieved by the proposed design. This attempt can be considered as the first to fabricate solar cells from highly-pure nanopowders of two different semiconductors.
In the present study, cytogenetic and molecular techniques were conducted to detect the chromosomal aneuploidy and the involvement of N and H genes in squamous larynx carcinoma cell line Hep-2.Our results showed that numerical and structural abnormalities were involved in larynx cancer Hep-2.The total number of chromosomes ranging from tripolyploidy in passage187to more than that in passage207.The more frequent chromosomes involved in numerical aberrations were chromosomes1,7,16,17 and 18. Structural chromosomal aberrations were also detected.Deletion of short arm was detected in chromosome 1(del 1p) and the long arm of chromosome 1(del 1q)and 6(del 6q).Gaining on short arms were also recorded in chromosomes 3(3p+) and 12(12p+).At the mole
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