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.
In many industries especially oil companies in Iraq consumed large quantities of water which will produce oil-contaminated water which can cause major pollution in agricultural lands and rivers. The aim of the present work is to enhance the efficiency of dispersed air flotation technique by using highly effective and cost-efficient coagulant to treating gas oil emulsion. The experimental work was carried out using bubble column made of Perspex glass (5cm I.D, 120cm height). A liquid was at depth of 60cm. Different dosage of sawdust +bentonite at ratio 2:1 (0.5+ 0.25; 1+ 0.5 and 2+1) gm and alum at concentration (10,20and30mg/l) at different pH ( 4 and 7) were used to determine optimum dosages of coagulant. Jar test exper
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Microalgae have been increasingly used for wastewater treatment due to their capacity to assimilate nutrients. Samples of wastewater were taken from the Erbil wastewater channel near Dhahibha village in northern Iraq. The microalga Coelastrella sp. was used in three doses (0.2, 1, and 2g. l-1) in this experiment for 21 days, samples were periodically (every 3 days) analyzed for physicochemical parameters such as pH, EC, Phosphate, Nitrate, and BOD5, in addition to, Chlorophyll a concentration. Results showed that the highest dose 2g.l-1 was the most effective dose for removing nutrients, confirmed by significant differences (p≤0.05) between all doses. The highest removal percentage was
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ABSTRACT:. The Lower Cretaceous Zubair formation is comprised of sandstones intercalated with shale sequences. The main challenges that were encountered while drilling into this formation included severe wellbore instability-related issues across the weaker formations overlaying the reservoir section (pay zone). These issues have a significant impact on well costs and timeline. In this paper, a comprehensive geomechanical study was carried out to understand the causes of the wellbore failure and to improve drilling design and drilling performance on further development wells in the field. Failure criteria known as Mogi-Coulomb was used to determine an operating mud weight window required for safe drilling. The accuracy of the geomechanical
... Show MoreThe study included the collection of 75 bronchial wash samples from patients suspected to have lung cancer. These samples were subjected to a diagnostic cytological study to detect the dominant type of lung cancer. It was noticed that 33 patients proved to have a lung cancer out of 75 (44%) of these, 19 cases (57.6%)were diagnosed having Squamus cell carcinoma,7cases (21.21%) showed Adenocarcinoma ,6 cases (18.18%) were having small cell carcinoma while only one case (3.03%)was large cell carcinoma .Nearly 70% of cases were correlated with smokers .Bacteria were isolated from 53 patients in which 33 isolates were associated with the cancer cases while 20 of them from non infected patients. By using different morphological ,biochemical test
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Toxic dyes are commonly discharged into waste waters and dyes are extensively used in the textile industry so it is necessary to find out efficient and eco-friendly method for treating waste waters resulting from industrial effluences. To achieve this aim the fungus Trichoderma sp. is employed into two lines: first line was self – immobilized fungal pellets in (Czapek – Dox medium) to adsorbs two dyes crystal violet, congo red by concentrations 0.01, 0.02, 0.03, 0.04, 0.05, 0.06 mg/L to both dyes, PH 2, room temperature with shaker in ( hrs.2,hrs.4,hrs.24) , by Uv- Visible spectrum . the removal efficiency of 0.05 mg/L crystal violet by Trichoderma sp was 96%. but there was no remova
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Phenomena of an abnormal genitalia was among some specimens of Cicindela aulica Dej.
Collected from Iraq. The fore tarsi of male were asymmetrical having its basal three segments
dilated and clothed beneath with fine bristles as in normal male. While those of the right leg
were found simple as in normal females. Dissection of the genialia of these specimens
showed that they were of two types of both male and female structures.
LK Abood, RA Ali, M Maliki, International Journal of Science and Research, 2015 - Cited by 2
A new species, Hottentotta pooyani sp. nov. is described and illustrated using two female specimens collected from the Khuzestan Province in Iran. This species is compared with the closely species including H. khoozestanus Navidpour, Kovařík, Soleglad & Fet, 2008; H. pellucidus Lowe, 2010 and H. saxinatans Lowe, 2010 that were previously described in Iran and Oman.
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