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 monitor and analyze the visual and symbolic features, derived from the Omani heritage, and inspired by the concept of local identity, for a selection of contemporary Omani graphic art works, which represent a mixed, multi-category of research sample, comprising three levels: the works of professional Omani artists, and the works of young artists and specialized students, as well as the work of a number of active academics in the field of graphic theorizing and teaching. The sample is also divided - in terms of technical classification - into a category of works executed using traditional engraving and printing media, and a category of works executed using modern and digital graphic printing media.
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Many international companies have branches or agencies in Iraq and have large market shares, and this has contributed to the increase in the import of these companies' products. This process created an additional problem of handling a large number of expired or defective products. There are many methods that contribute significantly to reducing these problems. The most prominent of these is the activation of reverse logistics options in Iraq by international and local companies. This research aims to conduct a survey of the reverse logistics options approved by some local and international companies and compare between them. This paper found that Amaron, Hitachi, Apple, Samsung and Huawei have limited options for reverse logistics and that
... Show MoreBackground: Semen contamination is a detrimental factor in decreasing fertility. Seasonal changes may affect the contamination, too. Objectives: This study was designed to detect semen contamination in ovine and caprine during different seasons. Methods: Six fully mature male sheep and goats were subjected to electro-ejaculator collection twice monthly from February 1, 2022, to January 31, 2023 (Spring, February 1, 2022-April 30, 2022; Summer, May 1, 2022, July 31, 2022; Autumn August 1, 2022, October 31, 2022; Winter November 1, 2022, January 31, 2023), for studying the seasonal effect. A total of 288 semen samples were collected from both species (36 samples from each per season). All samples were subjected to bacterial isolatio
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Buffering of Local anaesthesia (LA) has been suggested as a mechanism to improve injection comfort and hasten the onset of anaesthesia. Aim This study aimed to evaluate the effectiveness of buffered LA in the extraction of maxillary premolars and molars. Materials and Methods This randomized controlled study included 100 patients who were indicated for extraction of maxillary posterior teeth, they were randomly divided into two groups; a study group that received infiltration of buffered 2% lidocaine hydrochloride with 1:80,000 epinephrine LA, and a control group that received non-buffered 2% lidocaine hydrochloride with 1:80,000 epinephrine LA. The buffering was performed using the Onset® LA buffering system (Onpharma®). The outcome va
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60 patients diagnosed as having urticaria were included in the study ; 30 patients were effected with acute urticaria and 30 patients were affected with chronic urticaria. In addition, 30 healthy adult volunteers were selected as control group .The patients and control groups sera were examined with enzyme linked immunosorbent assay ( ELISA) to detect total level IgE and radial immunodiffusion (RID) to detect levels of IgG , IgA and IgM . The total level of IgE in acute urticaria ( 1.45±0.13) IU/mL and chronic urticaria (2.12 ± 0.10) IU/mL patients were significantly higher than the control groups ( 0.85 ± 0.10)IU/mL (p<0.05). The level of IgG in acute urticaria ( 12.5± 0.42) g/L and chronic (13.16±0.40) g/L patients , IgA in acute (2.
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The development of better tools for diagnosis and more accurate prognosis of cancer includes the search for biomarkers; molecules whose presence, absence or change in quantity or structure is associated with a particular tumour or prognosis/therapeutic outcome. While biomarkers need not be functionally relevant, if cell survival, then they could also provide new targets for therapeutic drugs. In recent years attention has been applied to a group of proteins known as cancer testis antigens (CT antigens) [1]. These proteins are products of genes whose expression was normally confined to the testis, yet they are expressed in tumour cells. CT genes are bound to serve a wide array of roles in the testes, which have many highly differentiated cel
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Echinococcosis is a zoonotic disease caused by the larval stage of the tapeworm Echinococcus granulosus. This disease is an important public health and a significant economic issue in Iraq, where the lungs and livers are the popular places of infection. The aim of the current study focused on using the molecular techniques in the detection of an E. granulosus strain that causes cystic echinococcosis to human, sheep and cattle in Thi-Qar province, Iraq. In the current study, thirty isolates of E. granulosus were collected from 10 human hydatid cysts through surgery done at Al-Hussein Imam Teaching Hospital in Thi-Qar province and 10 sheep with 10 cattle hydatid cysts were obtained from the slaughterhouse in Thi-
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