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.
Among a collection of leafhoppers from Erbil Province in Kurdistan/Iraq, a new species of the genus Arboridia Zakhvatkin, 1946 was designated and described here as a new species to the science. The erection of this species was mainly built on the external characters included the male genitalia. Sites and dates of collections so as the host-plants were verified.
Abstract:
In light of globalization and internationalization of financial markets, issues arising from financial crises have become increasingly serious and fundamental, creating a lot of debate among experts around the world. So, many studies have attempted to investigate what measures can be taken to detect and prevent crises before they devastate the economies.
Therefore, this paper examines the Effectiveness of the Monetary Policy (MP) to Avoid, Reduce or Treat the Financial Crisis in Malaysia. Scholars have yet to agree on the issue
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Artichoke (Cynara scolymus L.) is a nutritious vegetable that grown all over the world. It is a promising herbal plant, rich in bioactive components. It is considered as medicinal plant due to its nutritional and phytochemical composition, especially high proportion of phenolic compounds. The primary aim of this study was to achieve chemical profile analyses of artichoke for different phytochemcials, especially Scolymoside and Cynaroside. Methanolic crude was extracted from Artichoke leaves by rotary evaporator and separated by column chromatography. The fractions monitored by Thin Layer Chromatography (TLC), and identified in High-Pressure Liquid Chroma
... Show MoreIraqi crude Atmospheric residual fraction supplied from al-Dura refinery was treated to remove metals contaminants by solvent extraction method, with various hydrocarbon solvents and concentrations. The extraction method using three different type solvent (n-hexane, n-heptane, and light naphtha) were found to be effective for removal of oil-soluble metals from heavy atmospheric residual fraction. Different solvents with using three different hydrocarbon solvents (n-hexane, n-heptane, and light naphtha) .different variables were studied solvent/oil ratios (4/1, 8/1, 10/1, 12/1, and 15/1), different intervals of perceptual (15, 30-60, 90 and 120 min) and different temperature (30, 45, 60 and 90 °C) were used. The metals removal percent we
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Mammary tumors (CMT) in dogs in Iraq may be induced by carcinogenic war ordnance. In our study, 10 virgin un- spayed military/pet bitches aged 5-15 years presented with abnormal masses in the abdomen with painful oedema, swelling, anorexia, weight loss, weakness and mild fever. Examination of regional lymph nodes and thoracic radiography confirmed metastasis. Tumors were excised and determined to be mostly adenocarcinomas involving multiple glands, solid in texture, 5-15 cm in size, mostly in the inguinal mammary glands at stage T3: >5 cm. Microscopy confirmed presence of adenocarcinoma in 8 dogs and solid carcinoma in 2 with half of tumors being grade III. Tumors had pleomorphic hyperchromatic cell nuclei in stroma, epithelial cells of duc
... Show MoreIA Ali, FK Emran, DF Salloom, Annals of the Romanian Society for Cell Biology, 2021
Mastitis is an udder tissue inflammation which has infected various species of animals. It happens through several types of pathogenic bacteria, particularly Streptococcus agalactiae. GBS is a leading cause of cow mastitis. In our sample, 9.52% of Streptococcus agalactiae were isolated which were collected from bovine mastic milk and identified by biochemical tests such as catalase, oxidase, Production of indole, fermentation of sugar, an examination of antibiotic sensitivity, CAMP test and group kits of Lancefield. The results showed that all Streptococcus agalactiae isolate was diagnosed by CAMP test by the appearance of the arrowhead in blood agar and by the appearance of visible agglutination on a card in the serological grouping kit of
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