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.
Four metal complexes of Schi base ligand were prepared. To characterize both the ligand and the metal complexes, various techniques were employed, including elemental analysis, FT-IR spectroscopy, UV-Vis spectroscopy, molar conductivity measurements, magnetic moment determination, melting point, mass spectroscopy, 1H-NMR and 13C-NMR spectroscopy. The results showed that the metal complexes formed have the formula [M(L)2Cl2]Cl(n).H2O, where LD Schi base ligand and MDCr(III), Rh(III), Pt(IV) and Au(III), n D 1, 2. Based on spectroscopic analysis, coordination with metal ions involves the ’N’ donor atom of N atoms of the imine and the N-H amid group, and four complexes are suggested to have a six-coordinated octahedral structure. Molar c
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This study is designed to isolate and molecular identification of C. gattii, C. gattii is pathogenic yeast and effect immunocomposed and immunocompetent, Methods: collect 50 samples from eucalyptus leaves. The collection time was extended from November 2021 to February 2022 and then culture at SDA, Cryptococcus Differential Agar esculin agar and Eucalyptus leaves agar, Brain heart infusion agar with methyldopa and Brain heart infusion agar with methyldopa media, biochemical test including urease test, and then confirm identification by molecular identification by PCR technique sequencing and genetic analysis. The results showed that 4 swaps taken from eucalyptus leaves included cryptococcus neoformans. This study indicated that the virulenc
... Show MoreThe cuneiform texts considered one of the most important resources which play an important role help us to know how was the economic , politic and religious manner and every thing have relation of the history of Mesopotamia which contain many of information that enrich our knowldgement every side of Mesopotamia civilization since using writing as away to register the science and human knowldgement . The city of Sippar (Abu –Habba ) was one of most important city of ancient Mesopotamia which mentioned as one of first five Mesopotamian cities found before floating and it exposed through excavating rich one of them tablet which some of it dated to old Babylon period . The king Ammi-Saduqa has a big share of tablet we study number of thi
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This research has been prepared to isolate and diagnose one of the most important vegetable oils from the plant medical clove is the famous with Alaeugenol oil and used in many pharmaceuticals were the isolation process using a technique ultrasonic extraction and distillation technology simple
Objective of the study was to rate and analyze the physical fitness of female students from Iraq and Russia to find similarities, differences and regularities. Sampled for the study were 44 female non-sporting students: 20 students from Pedagogical University in Bagdad; and 24 students from Russian State University of Physical Education, Sport, Youth and Tourism in Moscow. The study was designed to obtain the joints flexibility, coordination, speed-strength and static endurance rates. The Iraqi students were tested with higher coordination abilities than their Russian peers, whilst the latter were ranked higher in the speed-strength, shoulder joints and spine flexibility tests, albeit lower than their Iraqi peers in the hip flexibility test
... Show MoreBiosorption of cadmium ions from simulated wastewater using rice husk was studied with initial concentration of 25 mg/l. Equilibrium isotherm was studied using Langmuir, Freundlich, BET and Timken models. The results show that the Freundlich isotherm is the best fit model to describe this process with high determination coefficient equals to 0.983. There was a good compliance between the experimental and theoretical results. Highest removal efficiency 97% was obtained at 2.5g of adsorbent, pH 6 and contact time 100 min.
The possibility of using zero-valent iron as permeable reactive barrier in removing lead from a contaminated groundwater was investigated. In the batch tests, the effects of many parameters such as contact time between adsorbate and adsorbent (0-240 min), initial pH of the solution (4-8), sorbent dosage (1-12 g/100 mL), initial metal concentration (50-250 mg/L), and agitation speed
(0-250 rpm) were studied. The results proved that the best values of these parameters achieve the maximum removal efficiency of Pb+2 (=97%) were 2 hr, 5, 5 g/100 mL, 50 mg/L and 200 rpm respectively. The sorption data of Pb+2 ions on the zero-valent iron have been performed well by Langmuir isotherm model in compared with Freundlich model under the studied
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There are numerous ways to define the traits of influential professors, including behavior (such as warmth, civility, and clarity), knowledge (of subject matter and students), and beliefs, among many others. According to research; influential teaching can also defined as the capacity to improve student achievement. This; is only one way to express effectiveness, as stated. However; the preferred definition of high-quality teaching in the United States and many other countries is teacher effects on student achievement. Therefore; this study aimed to determine the characteristics of influential professors in Online Teaching at the English department. This study; believed to be significant for college professors who will show them how
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