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 study included the extraction of volatile oil from Mentha piperita which was 1.3 % in the leaves and flowers . Volatile oil of the Mentha piperita leaves had special aromatic odour, pale yellow color, slightly pungent taste . The specific gravity and refractive index were (0.9794) and ( 1.464) respectively. The inhibition activity of the Mentha piperita Volatile oil extracts were studied on some pathogenic microorganisms like Staphylococcus aureus, Salmonella typhi, Escherichia coli, Proteus sp, and Klebsiella pneumoniae . The result showed that the volatile oil had an inhibition effect on the growth of all microorganisms, and it gave the higher inhibition effect on the growth of S. aureus in which the inhibition zone reached to 2
... Show MoreThis research presents a study for precipitating phosphorus (as phosphate ion) from simulated wastewater (5ppm initial concentration of phosphorus) using calcium hydroxide Ca(OH)2 solution. The removal of phosphorus by Ca (OH)2 solution is expected to be very effective since the chemical reaction is of acid-base type but Ca(OH)2 forms complex compound with phosphate ions called. Hydroxyapatite Ca5 (PO4)3OH. hydroxyapatite is slightly soluble in water. This research was directed towards sustainable elements as phosphorus. Kinetics of the dissolution reaction of hydroxyapatite was investigated to find the best factors to recover phosphorus. The effect of con
... Show MoreNeonatal sepsis refers to the bacterial bloodstream infections of the newborn during the neonatal period as usually the first twenty-eight days of life. The current study was done in the laboratories of AL-Batool Teaching Hospital for Gynecology and Pediatrics in Baqubah, Diyala Governorate, including 140 blood specimens collected from the neonates admitted to the hospital with suspected sepsis, the ages of the both groups was ranged from 1 day to 28 days. Out of the total cultured samples, 32.14% (45 of 140) were positive and 67.86% (95 of 140) were negative blood culture. 45 of 140 samples were negative to the blood culture chosen as control group. The results showed highest isolates were Coagulase Negative Staphylococcus (CoNS) 19 (42.2%
... Show MoreA total of 589 fishes, belonging to 23 species were collected from eight different localities
in north and mid Iraq during 1993. The parasitological inspection of such fishes revealed the
presence of 59 parasite species and two fungi. Among such parasites, five monogenetic
trematodes were recorded on the gills of some fishes for the first time in Iraq. These
included:- Ancyrocephalus vanbenedenii on Liza abu from Tigris river at Al-Zaafaraniya,
south of Baghdad; Dactylogyrus anchoratus on Cyprinus carpio from Tigris river at Al –
Zaafaranya D. minutus on C. carpio from both Tigris river at Al-Zaafaraniya and Euphrates
river at Al-Qadisiya dam lake; Discocotyle sagittata on L. abu from both the drainage system
at
The present work describes the adsorption of Ba2+ and Mg2+ions from aqueous solutions by activated alumina in single and binary system using batch adsorption. The effect of different parameters such as amount of alumina, concentration of metal ions, pH of solution, contact time and agitation speed on the adsorption process was studied. The optimum adsorbent dosage was found to be 0.5 g and 1.5 g for removal of Ba2+ and Mg2+, respectively. The optimum pH, contact time and agitation speed, were found to be pH 6, 2h and 300 rpm, respectively, for removal of both metal ions. The equilibrium data were analyzed by Langmuir and Freundlich isotherm models and the data fitted well to both isotherm modes as indicated by higher correlation of deter
... Show MoreBackground: Insertion sequence is a short DNA sequence encode for proteins implicated in the transposition activity. Transposase catalyzes the enzymatic reaction allowing the insertion sequence to +9*lo2 move. ;qqa;.
Objective: To study the sequencing of transposase gene, tnp, IS1216V of S. aureus isolated from food and then compared with that documented in National Center for Biotechnology Information (NCBI).
Methods: Food samples of animal
... Show MoreThe first aim of the present study was performed to assay the activity of arginase in sera of women with uterine fibroid.. This study consisted of(50) women with uterine fibroid as patient's group and (30) healthy women as control group. The age ranged between (30-55) years for the two groups. The results showed that highly significant increas (P< 0.0001) in the arginase activity in sera of women with uterine fibroid (7.99± 0.23) I.U/L is found when compared with healthy group (0.52±0.02) I.U/L. The second aim was performed to isolate arginase from sera of women with uterine fibroids. The purification is done by addition of ammonium sulfate, dialysis, gel filtration chromatography by using sephadex G-50 and ion exchange chromatography by
... Show MoreHCl is separated from HCl –H2SO4 solution by membrane distillation process(MD). The flat –sheet membranes made from polyvinylidene fluoride (PVDF) and polypropylene (pp.). Plate and frame these types of membrane where used in the process. The feed is a mixture of HCl and H2SO4 acids compositions depended on metals treated object.HCl concentration increased in the permeate during the process but sulfuric acid increased gradually in the feed .During the concentration of solution acids concentrations in the feed at the beginning were 50 g/dm3 of sulfuric acid and 50 g/dm3 of hydrochloric acid at 333K feed temperature the permeate flux was 71 dm
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