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.
Nanosilica was extracted from rice husk, which was locally collected from the Iraqi mill at Al-Mishikhab district in Najaf Governorate, Iraq. The precipitation method was used to prepared Nanosilica powder from rice husk ash, after treating it thermally at 700°C, followed by dissolving the silica in the alkaline solution and getting a sodium silicate solution. Two samples of the final solution were collected to study the effect of filtration on the purity of the sample by X-ray fluorescence spectrometry (XRF). The result shows that the filtered samples have purity above while the non-filtered sample purity was around The structure analysis investigated by the X-ray diffraction (XRD), found that the Nanosilica powder has an amorphous
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My research tagged [the lights of the statement in the first part of the Koran] came to show that the dear book was developed according to a precise linguistic system is not increased by a word or letter or movement - Aldmh and Kira and the hole - and does not lack anything of it except with the wisdom required by the meanings of the Koranic text or Sura generally . The Koran does not come falsehood from his hands or from behind it is infallible and preserved; because it was revealed by the sage Hamid Hamid ﭽﮗ ﮘ ﮙ ﮚ ﮛ ﮜ ﮝ ﮞ ﭼ [stone]. The Qur'an is safe from any verbal or moral dominance and dominates all the heavenly books and exists at any time and place that speaks the truth ﭽ ﯛ ﯜ ﯝ ﯞ ﯟ ﯹ ﯺﭼ [The
... Show MoreMicroorganisms have an active role in biotechnology for example yeasts, especially in some genus like Saccharomyces, Pichia, and Candida. C.tropicalis one of the most important species of Candida and despite it is one of the causative agents of candidiasis but it has a major role in the production of many chemical compounds. C.tropicalis in the previous study was isolated from sheep dung and morphologically and molecularly classified the result of sequencing was elucidate 100% similarity between the studied isolate and other isolates inserted in DNA Data Bank of Japan DDBJ, physiologically this isolate tolerated 6% ethanol concentration in broth media with the ability to the pro
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Four samples were collected from the wastewater of State Battery Manufacturing Company (SBMC); Babylon 2 factory in AL-Waziriya district, as triplicates. Physical and chemical measurements were carried out such as temperature, pH, Lead concentrations and their ranges were: (19.5-34.5) °C, (6.1-6.4) and (4.5-6.5) mg/L, respectively. Six dominant Bacillus spp. isolates were isolated from these samples; namely, Bacillus subtilis N1, Bacillus subtilis N2, Bacillus subtilis N3, Bacillus cereus N4, Bacillus cereus N5 , Bacillus cereus N6. These isolates were capable of removing Lead from aqueous solutions in a capacity reached 27.6 ± 1.4, 10.1 ± 1.7, 74.5 ± 0.7, 8.93 ± 2.8, 8.1 ± 3.5, 1.6± 0.7 mg/L, respectively. Whereas cell walls,
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Background: Birth defects are the leading cause of both neonatal and post-neonatal deaths, as an estimated 240,000 neonates die in their first month of life worldwide each year. In Iraq, local studies have shown varying frequencies and types of congenital anomalies. Objectives: To provide new insight into the incidence and types of congenital anomalies and to explore their possible risk factors in Baghdad City. Methods: In this cross-sectional observational study, a total of 2007 neonates were enrolled from six hospitals in Baghdad during the period extending between September and December 2020. Hospital records and personal interviews were used for data collection. These data included the neonates' demographic and clinical characte
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The lead has adverse effects in contamination the aquatic environment, for this reason, a laboratory simulation was conducted using kaolinite collected from the Ga’ara Formation at western Iraq to be considered as a natural sorbent material that can be addressed Pb2+ from the aqueous environments. The Energy-Dispersive X-ray Spectroscopy and atomic absorption spectroscopy clarifying very fine grains and pure phase with a very little quantity of quartz and has a number of active sites for adsorption. The sorption of kaolinite for the Pb2+ has been carefully tested by several designed laboratory experiments. Five lead solutions of different concentrations (25, 50, 75, 100 and 125 ppm) were tested under different values of pH (1.3-9)
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Urine specimen (253 samples) had been collected from urinary tract infections. The study showed that Proteus mirabilis was responsible for (11.85%) of the urinary tract infections. Also, the study had declared that the ratio of isolation of this bacterium from women was (7.51%) and it was higher than the ratio in both men and children which ranged (1.58%) and (2.76%) respectively. Morphological and biochemical studies had been applied to characterize this bacterium as well as other kinds of microorganisms that were isolated from urinary tract infection in this study. The study deals with typing methods such as using biotyping and typing according to Dienes phenomenon beside the succestibility to antibiotics. The results had shown that the s
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The present work aims to study the possibility of utilization a forward osmosis desalination process as an alternative method to extract water from brine solution rejected from reverse osmosis process.
Experiments conducted in a laboratory–scale forward osmosis (FO) unit in cross flow flat sheet membrane cell yielded water flux ranging from (0.0315 to 0.56 L/m2 .min) when using CTA membrane,and ranging from (0.419 to 2.785 L/m2 .min) for PA membrane under 0.4 bar. Two possible membrane orientations were tested. Sodium chloride with high concentrations was used as draw solution solute. The effect of membrane orientation on internal concentration polarization (ICP) was studied. Two regimes of ICP; dilutive and concentrative were desc
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