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.
Autorías: Omar Saeed Sabbar, Ali Mousa Jawad, Maher Amer Jabbar. Localización: Revista iberoamericana de psicología del ejercicio y el deporte. Nº. 3, 2023. Artículo de Revista en Dialnet.
Blastocystis is a ubiquitous human and animal protozoa that inhabit the gastrointestinal tract. Metronidazole is considered the standard drug for the treatment of Blastocystis infection; however, there is growing evidence of treatment failure, hazardous side effects, and appearance of strains resistant to metronidazole. In the last era, many studies have been implicated in the quest for new treatments for Blastocystis infection, especially natural products. Attention has been focused on the effect of Amygdalin (B17) and pumpkin seed on eradicating parasitic infections. The current work was built up to explore the in vitro efficacy of two natural compounds, Amygdalin (B17) and pumpkin seeds against
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This study describes the preparation of a new bidentate Schiff base derived from the condensation of Isatin-3-hydrazone with 2-acetylthiophene and the preparation of new series of complexes with a good yield. The prepared ligand was characterized by IR, UV-Vis, C.H.N.S elemental analysis, 1H and 13C NMR, LC-Mass spectroscopy, and physical measurements. Its complexes were analyzed by C.H.N.S elemental analyses, UV-Vis., FTIR, NMR, LC-Mass Spectra, atomic absorption spectroscopy, magnetic susceptibility, and conductivity measurements The results from spectroscopy and measurement studies showed that the ligand coordinated to the metal ion as a bidentate ligand via oxygen and nitrogen, forming an octahedral geometry around it. In vitro antimicr
... Show MoreAdhesion (type 1 fimbriae) and host defense avoidance mechanisms (capsule or lipopolysaccharide) have been shown to be prevalent in Escherichia coli isolates associated with urinary tract infections. In this work, 50 uropathogenic Escherichia coli (UPEC) isolated from children with urinary tract infections were genotypically characterized by polymerase chain reaction (PCR) assay. We used two genes; fimH and kpsMTII, both of them previously identified in uropathogenic E.coli (UPEC) isolates. The PCR assay results identified fimH (90.0)% and kpsMTII (72.0)% isolates. In the present study, was also demonstrated that these genes may be included in both or one of them within a single isolate.
This paper compare the accurecy of HF propagation prediction programs for HF circuits links between Iraq and different points world wide during August 2018 when solar cycle 24 (start 2009 end 2020) is at minimun activity and also find out the best communication mode used. The prediction programs like Voice of America Coverage Analysis Program (VOACAP) and ITU Recommendation RS 533 (REC533 ) had been used to generat HF circuit link parameters like Maximum Usable Frequency ( MUF) and Frequency of Transsmision (FOT) .Depending on the predicted parameters (data) , real radio contacts had been done using a radio transceiver from Icom model IC 7100 with 100W RF
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A multistep synthesis was established for the preparation of a new vanillic acid-1, 2, 4-1triazole-3-thiol conjugate (
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Abstract
The Purpose of This Research is The Main Factors In out Comes Phenomena From Primary School Which in Creased in Lost Period in Iraq And to Find Solutions to The This Problem.
In Order to Achieve Al The Aim The Research Choose a Systematic Random Sample of School Records For Students in Some Primary Schools in Karkh and Rusafa and Year of Study (2010-2015) and Size (40) Samples, included (16) Variable , Collected in Form Prepared by The Research As a Way to Analyze The Data.
Remember to Summarize The (6) Main components Pay a Student to Drop out of Primary Schools in The Province of Baghdad are Arranged As Follows:
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Synthesis, characterization and pharmaceutical studies of schiff base from 2-pyrrolidinone derivative and imidazole-2-carboxaldehyde and corresponding complexes with Metal (||)
This study sought to investigate the impacts of big data, artificial intelligence (AI), and business intelligence (BI) on Firms' e-learning and business performance at Jordanian telecommunications industry. After the samples were checked, a total of 269 were collected. All of the information gathered throughout the investigation was analyzed using the PLS software. The results show a network of interconnections can improve both e-learning and corporate effectiveness. This research concluded that the integration of big data, AI, and BI has a positive impact on e-learning infrastructure development and organizational efficiency. The findings indicate that big data has a positive and direct impact on business performance, including Big
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