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.
Background: Verrucous carcinoma (VSCC) is considered as a rare well differentiated carcinoma variant of SCC with no metastatic potential. The aim of this study is to evaluate the Immunohistochemical expression MMP-2, VEGF and D2-40 expression in OSCC and VSCC. Materials and methods: Thirty formalin fixed paraffin embedded tissue blocks of OSCC and another twelve VSCC were collected and Four micrometer thick sections were cut from each block and mounted on positively charged slides and stained immunohistochemically with monoclonal antibodies to MMP-2, VEGF and D2-40. Results: there is no statistical difference between SCC and VSCC regarding the immunoexpression of MMP-2 and VEGF. While the lymphatic vessels density were higher in SCC than V
... Show More
This paper aims at the analytical level to know the security topics that were used with data journalism, and the expression methods used in the statements of the Security Media Cell, as well as to identify the means of clarification used in data journalism. About the Security Media Cell, and the methods preferred by the public in presenting press releases, especially determining the strength of the respondents' attitude towards the data issued by the Security Media Cell. On the Security Media Cell, while the field study included the distribution of a questionnaire to the public of Baghdad Governorate. The study reached several results, the most important of which is the interest of the security media cell in presenting its data in differ
... Show More
In today's world, the science of bioinformatics is developing rapidly, especially with regard to the analysis and study of biological networks. Scientists have used various nature-inspired algorithms to find protein complexes in protein-protein interaction (PPI) networks. These networks help scientists guess the molecular function of unknown proteins and show how cells work regularly. It is very common in PPI networks for a protein to participate in multiple functions and belong to many complexes, and as a result, complexes may overlap in the PPI networks. However, developing an efficient and reliable method to address the problem of detecting overlapping protein complexes remains a challenge since it is considered a complex and har
... Show More
Background: Diabetes mellitus is a chronic disease with an increasing prevalence worldwide and characterized by an increase in oxidative stress and inflammation. The most important factor that is responsible for oxidative stress and production of reactive oxygen species (ROS) is hyperglycemia. The major targets of ROS are proteins. The most common and widely used biomarker of severe oxidative protein damage is protein carbonyl content.
The study was designed to assess the serum level of protein carbonyl as a marker of protein oxidation in patients with type 2 diabetes mellitus and to evaluate the effect of age, body weight, waist circumference, diabetic control and disease duration on the level
... Show More
(4)
This study is concerned with the derivation of differential equation of motion for the free coupled vertical – torsional and lateral vibration of opened thin-walled curved beams. The curved beam to be considered in this study is of isotropic opened thin – walled (I) section with equal top and bottom flanges. The derivation depends on Hamilton's principle which required finding the potential and kinetic energy of the curved beam section due to internal stresses and all types of movements (Vertical,Torsional and Lateral) .The effect of restrained warping displacement is also considered in this study. Three differential equations are derived for vertical, torsional and lateral movement .and approximate solutions are developed by using the
... Show MoreWater is the basis of the existence of all kinds of life, so obtaining it with good quality represents a challenge to human existence and development especially in the desert and remote cities because these areas contain small populations and water purification requires great materials and huge amounts of fossil fuels resulting pollution of the environment. Cheap and environmentally friendly desalination methods have been done by using solar distillations. Passive solar stills have low yields, so in this research, the problem is overcome by connecting four heat pipes which are installed on the parabolic concentrator reflector with passive solar still to increase the temperature of hot water to more than 90°C, as a resul
... Show More
(3)
(2)
The present study aims to detection optimal conditions of production of amylase enzyme from isolate of B. subtillis A4. Nine carbonic sources were represented by starch, maltose, fructose, sucrose, glucose, arabinose, xylose, sorbitol and mannitol) at concentration of 1% for each source. It was found that the best was represented by starch carbonic, which showed higher activity and qualitative activity of 7.647 Unit/ ml and 461.56 Unit/ mg. Ten nitrogen sources were selected, including yeast extract, peptone, trypton, gelatin, urea and meat extract as organic sources Ammonium sulphate, Sodium nitrate, Potassium nitrate and Ammonium chloride as inorganic sources. These sources were added at aconcentration of 0.5% to the production medium. Th
... Show More
(2)
This work deals with preparation of zeolite 5A from Dewekhala kaolin clay in Al-Anbar region for drying and desulphurization of liquefied petroleum gas. The preparation of zeolite 5A includes treating kaolin clay with dilute hydrochloric acid 1N, treating metakaolin with NaOH solution to prepare 4A zeolite, ion exchange, and formation. For preparation of zeolite 4A, metakaolin treated at different temperatures (40, 60, 80, 90, and 100 °C) with different concentrations of sodium hydroxide solution (1, 2, 3, and 4 N) for 2 hours. The zeolite samples give the best relative crystallinity of zeolite prepared at 80 °C with NaOH concentration 3N (199%), and at 90 and 100°C with NaOH concentration solution 2N (184% and 189%, respectively). Ze
... Show More