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 research aims to use a new technology for industrial water concentrating that contains poisonous metals and recovery quantities from pure water. Therefore, the technology investigated is the forward osmosis process (FO). It is a new process that use membranes available commercial and this process distinguishes by its low cost compared to other process. Sodium chloride (NaCl) was used as draw solution to extract water from poisonous metals solution. The driving force in the FO process is provided by a different in osmotic pressure (concentration) across the membrane between the draw and poisonous metals solution sides. Experimental work was divided into three parts. The first part includes operating the forward osmosis process using T
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The members of the family of Eentrobacteriaceae harbour a gene cluster called polyketide synthase (pks) island. This cluster is responsible for the synthesis of the genotoxin colibactin that might have an important role in the induction of double-strand DNA breaks, leading to promote human colorectal cancer (CRC). Eleven out of the eighty eight isolates (12.5%) were pks+, distributed as 7 (8%) isolates of E. coli, 2 (2.25%) of K. pneumoniae and 2 (2.25%) of E. aerogenes. The cytotoxic effects of selected pks+ isolates (E. coli and E. aerogenes) on HeLa cells were represented by decreasing cell numbers and enlarged cell nuclei in comparison to the untreated cells. Cytological changes were observed when the infected HeLa cells culture
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Nine fish specimens of Thryssa setirostris (Broussonet, 1782) were collected from the Arabian Gulf, during the period from July 2015 to April 2016. Because of the scarcity of this fish and overlapping and ambiguous of its taxonomic characters with other Thryssa spp., a detailed taxonomic study was conducted. The present study includes the most important meristic and morphometric characteristics.
The mean of the total length of the specimens was 149.67 mm; dorsal fin consists of 12 rays, anal fin with 34-37 rays and pectoral fin with 12-13 rays; Gill rakers were 4 upper,1 medial an.10 lower. The most important character that isolates T. setiristis from
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Copper oxide (CuO) nanoparticles were synthesized through the thermal decomposition of a copper(II) Schiff-base complex. The complex was formed by reacting cupric acetate with a Schiff base in a 2:1 metal-to-ligand ratio. The Schiff base itself was synthesized via the condensation of benzidine and 2-hydroxybenzaldehyde in the presence of glacial acetic acid. This newly synthesized symmetric Schiff base served as the ligand for the Cu(II) metal ion complex. The ligand and its complex were characterized using several spectroscopic methods, including FTIR, UV-vis, 1H-NMR, 13C-NMR, CHNS, and AAS, along with TGA, molar conductivity and magnetic susceptibility measurements. The CuO nanoparticles were produced by thermally decomposing the
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The goal of current research is to identify the difficulties in the application of modern physics in the middle schools of the province of Baghdad schools from the perspective of teachers of physics trends, sample search of (127) teachers, Karkh Third Directorate, and use Researcher questionnaire data collection tool after applying it to teachers who have experience (5) years and more after confirmation of the validity and reliability of the scale (the tool) researcher has used the averages for the purpose of interpreting the results. the results showed that the difficulties have been in the order following: (difficulties related educational environment of modern trends of teaching, curriculum-related dif
... Show MoreThis study aims to investigate the academic leaders’ perceptions towards the degree of availability of the dimensions of organizational immunity systems at the University of Tabuk, as well as to reveal the statistically significant differences between the average responses of the study sample members about the degree of availability of these dimensions at their university due to the variables of (gender, leadership position, and college specialization). To achieve the objectives of the study, a descriptive survey method was used. The study population consisted of (200) male and female leaders who were academic leaders at the University of Tabuk. A questionnaire was used as a tool for collecting data, which its validity and reliability
... Show MoreThe new liganed Schiff base named [(E)-3-hydroxy-4-((3,4,5- trimethoxybenzylidene)amino) naphthalene-1- sulfonic acid] was synthesized from 3,4,5-trimethoxybenzyldehyde and 1-amino-2-aphthol-4- sulfonic acid in equal molar ratio. A series of new metal complexes' of the common molecular formulation [M(L)2(H2O)2].H2O are synthesized and characterized by IR, UV–Vis spectra, mass spectra, atomic absorption, elemental analyses, chloride content, magnetic susceptibility and conductivity measurements as well as thermo gravimetric analysis (TGA, DSC). Consistent with results of the magnetic and spectral studies, the advised geometrical structures for all of the prepared complexes have been octahedral formula
The study was aims to evaluate the antimicrobial acttvtty of petroleum ether extracts from leaves , seeds and root of Zygophyllum fabago , against several microorganisms including gram negative bacteria (Pseudomonas aeruginosa & Escherichia coli), gram positive bacteria (Staphylluwccus aureus & Bacillus subtilis), in addition to yeast (Candida albicans).
While the results of sensitivity of the microorganisms to words petroleum ether extracts showed different activity , petrolewn ether extract of seeds showed more antimicrobial
... 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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