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: Pathogenic bacteria responsible for the causation of many common diseases have been identified on currency notes. The present investigation was carried out on one hundred currency notes of all the denominations (50, 100, 250, 500 and 1000RY), obtained from different occupational mainly bus drivers, hawker street, vegetable vendor, restaurants and butchers and fish seller groups in Taiz city,Yemen. Identification and characterization revealed active participation of the following species of organisms in the ascending order of percentage as E. coli(50.28 %),Staphylococci aureus(14.04 %), Klebsiellaspp(4.39 %),proteus(4.39 %), salmonella(1.25 %), shigella(0.72 %), Coagulase negative staphylococcus(0.60 %), pseudomonas(0.50 %),
... Show MoreThis study was carried out to find out the effect of germination of broad beans and chickpeas seeds for different periods on the sensory properties of homus bethina and falafel. The results revealed that the studied properties were significantly different (P<0.05) in tenderness, flavor and overall acceptance as compared to control samples. While other properties such as appearance, body (texture), leavening and color did not showed significant differences.It was found that treatment B1 (100% germinated broad beans) varied significantly in tenderness in comparison with control samples.Treatment B3 (75% ordinary bread beans + 25% germinated broad beans) revealed significant differences (P<0.05) in both flavor and overall acceptance as compar
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Calendula officinalis L. (Asteraceae) known as marigold is known to have several pharmacological activities and used for the treatment of several diseases as measles, jaundice, constipation and several inflammations. Marigold flowers contain several chemical constituents mainly flavonoids, triterpenoids and essential oil. In this study marigold flowers cultivated in Iraq had been investigated for its flavonoids content. The study revealed the presence of quercetin and kaempferol glycosides and the absence of myricetin glycosides. The flowers were extracted with ethanol 70% fractionated with different solvent and the flavonoids were isolated by preparative HPLC. The isolated flavonoids were identified by measuring melting points, UV, IR,
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This study is about awareness of teaching explanation difficulties in the Islamic university from the lecturers point of view. It discussed the difficulties and the traditional teaching methods of explanation. The study concentrated on teaching Islamic studies in general and teaching explanation in specific and set difficulties so as to be treated.
The study is of three chapters, the first contains the difficulties in several aspects like the educational goals, text contents, teaching methods and styles, students, educational techniques, educational aids and evaluation, it addition to the lecturers of Islamic university colleges in 2009-2
... Show MoreThis investigation deals with the use of orange peel (OP) waste as adsorbent for removal of nitrate (NO3) from simulated wastewater. Orange peel prepared in two conditions dried at 60C° (OPD) and burning at 500 °C (OPB). The effect of pH: 2-10, contact time: 30- 180 min, sorbent weight: 0.5- 3.0 g were considered. The optimal pH value for NO3 adsorption was found to be 2.0 for both adsorbents. The equilibrium data were analyzed using Langmuir and Freundlich isotherm models. Freundlich model was found to fit the equilibrium data very well with high-correlation coefficient (R2). The adsorption kinetics was found to follow pseudo-second-order rate kinetic model, with a good correlation (R2
... Show MoreThe Catharanthus roseus plant was extracted and converted to nanoparticles in this work. The Soxhlet method was used to extract alkaloid compounds from the Catharanthus roseus plant and converted them to the nanoscale. Chitosan polymer was used as a linking material and converted to Chitosan nanoparticles (CSNPs). The extracted alkaloids were linked with Chitosan nanoparticles by maleic anhydride to get the final product (CSNPs-Linker-alkaloids). The pure Chitosan, Chitosan nanoparticles, and CSNPs-Linker-alkaloids were characterized by X-ray diffractometer, and Fourier Transform Infrared spectroscopy. X-ray results show that all samples have an orthorhombic structure with crystallite size in nanodimensions. FTIR spectra prove that
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Online examination is an integral and vital component of online learning. Student authentication is going to be widely seen when one of these major challenges within the online assessment. This study aims to investigate potential threats to student authentication in the online examinations. Adopting cheating in E-learning in a university of Iraq brings essential security issues for e-exam . In this document, these analysts suggested a model making use of a quantitative research style to confirm the suggested aspects and create this relationship between these. The major elements that might impact universities to adopt cheating electronics were declared as Educational methods, Organizational methods, Teaching methods, Technical meth
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The -multiple mixing ratios of γ-transitions from levels of populated in the are calculated in the present work by using the a2-ratio methods. We used the experimental coefficient (a2) for two γ-transitions from the same initial state, the statistical tensor, which is related to the a2-coefficient would be the same for the two transitions. This method was used in a previous work for pure transitions or which can be considered pure. In these cases the multiple mixing ratios for the second transition ( ) equal zero, but in our work we applied this method for mixed γ-transitions and then the multiple mixing ratio ( ) is known for one transition. Then we calculate the ( ) value and versareversa. The weight average of the -values calcu
... Show MoreThe problem of research is to identify the uses of Iraqi women for Facebook and the motives of these uses and the innovations that have been achieved from them, in view of the increasing role-played by social networking sites in the lives of individuals in general and women in particular.The research seeks to identify the following: Identify the extent to which Iraqi women use Facebook and the usage motives and innovations achieved by the interviewees When they using Facebook. This study belongs to the descriptive studies that are interested in monitoring the characteristics of a particular phenomenon to identify its characteristics and characteristics.This study adopted the survey methodology to test the hypotheses of the study and how
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