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.
This research concerns the fact of the school garden and how can we tackle this issue. The research starts with a question forwarded to those who are interested in the ministry of education matters; the question is that how is the extent of interest in the school garden project? And how it is activated in the Iraqi Educational Institution program? In addition, the analysis reveals the importance of the school garden since we, as teachers and students, suffer from seeing the view of the trashes, the wastes of school furniture, the stinking water and the weeds and how it is accumulated in the back yard of the school. To add fuel to the fire, the increase in number of the students; year after year. This increase leads to adding more classes
... Show MoreBackground: The aim of this study was to determine phototoxic effect of visible blue light on anaerobic periodontal pathogens namely Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis. Materials and methods: Strains of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis were isolated from pockets of systemically healthy patients aged between 35-55 years old with pocket depths of 5-6 mm, the bacteria cultured on special blood Agar plates solid media, then subjected to visible blue light emitted from commercially available light cure devise (LED curing light); that emits blue light (400-500nm) of 1000mw energy at different periods of time exposures, then the CFU of each plate was measured by direct colony count
... Show MoreMost companies use social media data for business. Sentiment analysis automatically gathers analyses and summarizes this type of data. Managing unstructured social media data is difficult. Noisy data is a challenge to sentiment analysis. Since over 50% of the sentiment analysis process is data pre-processing, processing big social media data is challenging too. If pre-processing is carried out correctly, data accuracy may improve. Also, sentiment analysis workflow is highly dependent. Because no pre-processing technique works well in all situations or with all data sources, choosing the most important ones is crucial. Prioritization is an excellent technique for choosing the most important ones. As one of many Multi-Criteria Decision Mak
... Show More
(2)
This Paper aims to plan the production of the electrical distribution converter (400 KV/11) for one month at Diyala Public Company and with more than one goal for the decision-maker in a fuzzy environment. The fuzzy demand was forecasting using the fuzzy time series model. The fuzzy lead time for raw materials involved in the production of the electrical distribution converter (400 KV/11) was addressed using the fuzzy inference matrix through the application of the matrix in Matlab, and since the decision-maker has more than one goal, so a mathematical model of goal programming was create, which aims to achieve two goals, the first is to reduce the total production costs of the electrical distribution converter (400 KV/11) and th
... Show More
Mixed ligand metal complexes of CrIII, FeIII,II, NiII and CuII have been synthesized using 5-chlorosalicylic acid (5-CSA) as a primary ligand and L-Valine (L-Val) as secondary ligand. The metal complexes have been characterized by elemental analysis, electrical conductance, magnetic susceptibility measurements and spectral studies. The electrical conductance studies of the complexes indicate their electrolytic nature. Magnetic susceptibility measurements revealed paramagnetic nature of the all complexes. Bonding
The research aims to know the relation between positive thinking and self academic motivation, trend towards teaching profession. So a sample of (344) male and female student of the morning study of education colleges from the two universities of Baghdad and al Mustansria for the academic year 2012-2013.
So three tools had been used to achieve the procedures of the research which are: the scale of positive thinking constracted by Abdal satar Ibrahim year 2010, the scale of self academicmotivation constracted by (Gottfried) and adapted by Hanan Hussan year 2010, the scale of teaching profession trend constracted by Enayat Zaki year 1974.
And by using the statistical means of person
... Show MoreThe heavy metals Cd, Cu, Fe, pb, and Zn were determined in dissolved and particulate phases of the water,in addition to exchangeable and residual phases of the sediment and in the selected organs of the fish Cyprinus carpio collected from the Euphrates River near Al-Nassiriya city center south of Iraq during the summer period / 2009 .Also sediment texture and total organic carbon(TOC) were measured. Analysis emploing a flam Atomic Absorption Spectrophotometers . The mean regional concentrations of the heavy metals in dissolved (µg/l) and particulate phases (µg/gm) dry weight were Cd (0.15,16.13) ,Cu (0.59,24.48) ,Fe (726,909.4) ,Pb (0.20, 49.95) and Zn (2.5,35.62) respectively,and those for exchangeable and residual phases of the
... Show More
The present study aims at identifying the effect of Renzulli model on achievement and holistic thinking among the fifth grade students in the Holy Quran and Islamic Education. The experimental method with partial control was used, and the sample was chosen randomly. The sample consisted of (62) students distributed into experimental and control groups. 164 behavioral goals were formulated based on Bloom's first three levels taxonomy (knowledge and comprehension, application, analysis and installation, and evaluation). The researchers designed a post-test to measure the achievement of students in the subject of Holy Quran and Islamic Education which consisted of (40) objective items and a measure of holistic thinking which consisted of (3
... Show More