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.
The present study investigates the characterization of silver nanoparticles (AgNPs) synthesized using Fusarium solani and their impact on tomato seed germination, plant growth, and disease resistance. A visible color change from yellow to dark smoky indicated the formation of AgNPs, while UV-visible spectrophotometry revealed an absorbance peak at 437 nm, confirming their presence. Atomic force microscopy analysis showed that the AgNPs ranged from 0 to 39.27 nm in size, with an average height of 5.772 nm, while scanning electron microscopy highlighted their diverse surface morphology. The application of AgNPs and mycorrhizal fungi significantly improved tomato seed germination rates, plant height, and dry weight compared to untreate
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The current research seeks to know the difficulties in memorizing literary texts with fourth grade students from teachers’ point of view of and students
The research contains a community of (10,870) students from schools affiliated to the Directorate of Karkh, second and the number of schools for boys and girls (79) School and the number of teachers (349) who are specialized in teaching Arabic language to fourth grade preparatory students and there was a section of a randomly sample of each of the students, teachers and schools .
Some actions which can be listed below:-The survey which was done by the researcher for the views of the two samples from teachers and students as the number of teachers covered in this research (20) and
Objectives: Six different Schiff bases were synthesized from ampicillin and amoxicillin with isatin, 5-bromoisatin, and 5-nitroisatin. Methods: Ampicillin and Amoxicillin are linked directly through their α-amino groups to the acyl side chain with isatin and isatin derivatives by nucleophilic addition using glacial acetic acid as a catalyst. Results: chemical structures of these Schiff bases were confirmed using FTIR, 1H NMR and elemental microanalysis. The antibacterial activity was evaluated by measuring minimum inhibitory concentration (MIC) values and showed various degrees of antibacterial activities when compared with parent drugs. Compounds 1a and 2b, which are the Schiff bases of ampicillin and amoxicillin with isatin, showed very
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The adsorption of Pb(II) ions onto bentonite and activated carbon was investigated. The effects of pH, initial adsorbent dosage, contact time and temperature were studied in batch experiments. The maximum adsorption capacities for bentonite and activated carbon were 0.0364 and 0.015 mg/mg, respectively. Thermodynamic parameters such as Gibbs free energy change, Enthalpy change and Entropy change have been calculated. These thermodynamic parameters indicated that the adsorption process was thermodynamically spontaneous under natural conditions and the adsorption was endothermic in nature. Experimental data were also tested in terms of adsorption kinetics, the results showed that the adsorption processes followed well pseudo second- order
... Show MoreIn this investigation a high density polyethylene (HDPE) was used as a substitute to polyvinylchloride in the production of lead acid battery separators. This has been achieved by preparing mixtures of different percentages of the feed materials which include a high density polyethylene (HDPE) locally produced, filler materials such as silica and oils such as dioctylphthalate (DOP) or paraffin which were added to the mixture to improve the final properties of the separator. The materials were compounded by two roll-mills under the same conditions. The following parameters are involved: &nb
... Show MoreHelminthiasis is a significant economic burden on grazing cattle. Increased resistance to currently available synthetic anthelmintics used to treat helminthiasis, and anthelmintic residues in meat and dairy products pose a significant worldwide health threat. These obstacles require the development of new anthelmintics capable of combating drug resistance while also exhibiting improved safety profiles. Rhynchosia cana (Fabaceae) is a herb that has historically been used as a worm expeller. To evaluate the phytochemical profile and explore the anti-oxidant and anthelmintic effects of different extracts of Rhynchosia cana (R. cana) by In silico and In vitro methods. Using standardised chemical t
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In this study, a proposed process for the utilization of hydrogen sulphide separated with other gases from omani natural gas for the production of sulphuric acid by wet sulphuric acid process (WSA) was studied. The processwas simulated at an acid gas feed flow of 5000 m3/hr using Aspen ONE- V7.1-HYSYS software. A sensitivity analysis was conducted to determine the optimum conditions for the operation of plant. This included primarily the threepacked bed reactors connected in series for the production of sulphur trioxidewhich represented the bottleneck of the process. The optimum feed temperature and catalyst bed volume for each reactor were estimated and then used in the simulation of the whole process for tw
... Show MoreThis note reported the first record of Spotted Flycatcher Muscicapa striata (Pallas, 1764) (Passeriformes, Muscicapidae) from the state of Odisha, India. This species was recorded from the north and western part of the country as well as from the Western Ghats, but this note reports the first record from the Eastern Ghats of India.
The current research aims to investigate the counseling services provided to gifted students in the Al-Baha region from their point of view, and to identify the differences in these views according to the students ’gender, educational stages, and the cities in which they study. The descriptive survey approach was followed in its relevance to the research objectives. Likewise, a random sample of gifted elementary, intermediate, and secondary school students was selected by the method of stratified sampling, as the sample reached (175) male and female students. The instrument of the research consisted of the questionnaire of counseling services presented to gifted students from their point of view prepared by researchers. The results sho
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