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.
TV drama has gone through many developmental stages until it reached and settled in the form of TV series of thirty episodes. Alongside the development of the TV technologies and the widespread of satellite channels, the form of the TV drama has changed and the series has consisted of two parts and then parts multiplied until they amounted to ten parts and more. This form of TV drama has become an artistic phenomenon, that once the series is displayed on one of the channels and achieves a noticeable success, its producers work to produce a second part of that series and so on. This form of TV drama has remained away from being researched.
This has urged the researcher to accomplish this study entitled (TV series of multiple parts fro
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The apricot plant was washed, dried, and powdered after harvesting to produce a fine powder that was used in water treatment. created an alcoholic extract from the apricot plant using ethanol, which was then analysed using GC-MS, Fourier transform infrared spectroscopy, and ultraviolet-visible spectroscopy to identify the active components. Zinc nanoparticles were created using an alcoholic extract. FTIR, UV-Vis, SEM, EDX, and TEM are used to characterize zinc nanoparticles. Using a continuous processing procedure, zinc nanoparticles with apricot extract and powder were employed to clean polluted water. Firstly, 2 g of zinc nanoparticles were used with 20 ml of polluted water, and the results were Tetra 44% and Levo 32%; after
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The availability of low- cost adsorbent namely Al-Khriet ( a substance found in the legs of Typha Domingensis) as an agricultural waste material, for the removal of lead and cadmium from aqueous solution was investigated. In the batch tests experimental parameters were studied, including adsorbent dosage between (0.2-1) g, initial metal ions concentration between (50-200) ppm (single and binary) and contact time (1/2-6) h. The removal percentage of each ion onto Al-Khriet reached equilibrium in about 4 hours. The highest adsorption capacity was for lead (96%) while for cadmium it was (90%) with 50 ppm ions concentration, 1 g dosage of adsorbent and pH 5.5. Adsorption capacity in the binary mixture were reduce at about 8% for lead a
... Show MoreThe research talks about the most important challenges facing Muslim youth of their ideological, social and economic types, and the youth is facing several problems, the most important of which are the intellectual and social invasion to which the Islamic nation has been exposed and ways to address them from a Quranic perspective and find solutions to these problems and these challenges in accordance with Islamic Sharia and the texts of the Holy Quran. From three topics and several demands, during which the researcher tried to find solutions to each challenge through the verses of the Noble Qur’an.
The aim of this study is to propose reliable equations to estimate the in-situ concrete compressive strength from the non-destructive test. Three equations were proposed: the first equation considers the number of rebound hummer only, the second equation consider the ultrasonic pulse velocity only, and the third equation combines the number of rebound hummer and the ultrasonic pulse velocity. The proposed equations were derived from non-linear regression analysis and they were calibrated with the test results of 372 concrete specimens compiled from the literature. The performance of the proposed equations was tested by comparing their strength estimations with those of related existing equations from literature. Comparis
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This research consists of two parts, the first part concern with analyzing the collected data of BOD and COD values in discharge waste water from Al-Dora refinery during 2010 to find the relationship between these two variables The results indicates that there
is a high correlation between BOD and COD when using a natural logarithm model (0.86 ln(COD)) with correlation coefficient of 0.98. This relationship is useful in predicting the BOD value using the COD value. The second part includes analyzing collected data from the same site in order to find a relationsip between BOD and other parameters COD, Phenol(phe), Temperature(T), Oil, Sulphat(SO4),pH and Total dissolved solids( TDS) discharged from the refinery. The results indicated
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Several specimens of the avocet, Recurvirostra avocetta L. are found infected with
Himantocestus gigantivcus sp. nov. ( Cestoda , Diploposthidae) . This cestode is related to H.
blanksoni Ukoli 1965 but easily differentiated from it in having longer and wider strobila,
larger size of testes but lesser in number, cirrus situated in the middle of mature segment
histead of anterior third and slightly posterior to the middle in gravid segment instead of the
middle , ovary and vitelline gland are larger , and the uterus has more branches.